Increasing the Level of Peroxisome Proliferator-Activated Receptor Gamma

Increasing the level of Peroxisome Proliferator-Activated Receptor Gamma

Coactivator 1 α in podocytes results in collapsing glomerulopathy

Supplement Figure 1. Regulation of PGC1α expression in cultured podocytes.

Relative transcript levels of PGC1α in podocytes were measured by QRT PCR. Abbreviation: Tumor necrosis factor alpha (TNF), Vascular Endothelial Growth Factor A (VEGF-A), Lipopolysaccharides (LPS), Transforming Growth Factor Beta 1 (TGF-b). n=3 each group. *** q<0.001 compared to control, ANOVA post hoc test.

Supplement Figure 2. Restoring PGC1α in podocytes does not ameliorate albuminuria in DKD model.

PGC1α transgene expression was titrated by doxycycline chow amount, after intensive-dose titration, we found a two-fold PGC1α is tolerable to podocytes and above that is toxic. In a 16 week STZ-induced DKD model, non-diabetic Nefta-PGC1α did not have more albuminuria if PGC1α expression is controlled in this window, but we did not observe a protective effect of diabetic albuminuria in Nefta-PGC1α mice. * p<0.05 compared to control group, ANOVA post hoc test, n=4 each group.

Supplement Figure 3. qPCR analysis of cultured podocytes infected with control (GFP) or adenoviral PGC1α

(a) Notch pathway gene expression in control and PGC1α expressing cells. Abbreviation: Hes Related Family BHLH Transcription Factor YRPW Motif 1 (Hey1), Hes Family BHLH Transcription Factor 1 (Hes1), Jagged 1 (Jag1). (b) HIF pathway gene expression in control and PGC1α expressing cells. Abbreviation: Hypoxia Inducible Factor 1 Alpha Subunit (Hif1a), Hypoxia Inducible Factor 2 Alpha Subunit (Hif2a), Twist Family BHLH Transcription Factor 1 (twist), Lysyl Oxidase (Lox), Lactate Dehydrogenase A (Ldha), Vascular Endothelial Growth Factor A (Vegfa). (c) PDGF pathway gene expression in control and PGC1α expressing cells. Abbreviation: Platelet Derived Growth Factor (Pdgf). (d) Tgf-β pathway gene expression in control and PGC1α expressing cells. Abbreviation: Transforming Growth Factor Beta 1 (Tgf-b), SMAD Family Member 7 (Smad-7), Snail Family Transcriptional Repressor 1 (Snail1), Snail Family Transcriptional Repressor 2 (Snail2). *q<0.05 compared to control, unpaired t test with Bonferroni correction.

Supplement figure 4. Block fatty acid utilization does not attenuate PGC1α induced podocytes proliferation and de-differentiation in vitro.

p=ns p=ns

3 ad-GFP + DMSO ) n n i

t ad-GFP + etomoxir o i c s a

s 2 ad-Pgc1a + DMSO o e t r

p p=ns d

x ad-Pgc1a + etomoxir e e z

i l e a v i

t 1 m a r l o e r n (

c E o y n p m li n - c y C y S c

Cultured podocytes were treated by CPT-1 specific inhibitor etomoxir (40uM) or DMSO before ad-GFP or ad-Pgc1a infection. Etomoxir was not able to attenuate PGC1α induced proliferation (C-myc, cyclin E) and de-differentiation (Synpo). n=3 each group, t-test.

Supplement Figure 5. VEGF-a protein expression in Control and Nefta-PGC1α glomeruli

p=ns

1.5

1.0

0.5 VEGF expression

0.0 a Control Nefta-PGC1 Glomeruli from control and Nefta-PGC1α mice were isolated by dynabeads perfusion, VEGF-a protein level was analyzed by western blot and quantified. N=6 each group, t-test.

Supplement Figure 6. A trend of increased PGC1α and its downstream gene expression in human CGN.

a b

p = 0 .1 2 2 p = 0 .1 4 1 1 .5 3 d e A r d 1 1 e e F t r C 2 n e R G t 1 .0 e N n R c e f - A

c 1 o n - P

a n y i P t a i d i f s d e o

0 n e 0 .5 y m e t

i m t

2 s n 2 i n g

g -1 e o t l o l n i -2 0 .0 C o n tro l C G N C o n tro l C G N

c d p = 0 .0 1 6 p = 0 .0 0 4 4 3 9 d 1 e r d A e e B 5 3 r 2 t 1 2 e n t T e C n P c L e -

c C 1 S n -

f 2 n a o i

a o i d

y d t y e i 0 t e i s m s n

n e

2 1 t 2 e g n t g -1 i o o n l l i

-2 0 C o n tro l C G N C o n tro l C G N

e p = 0 .0 3 2 f p = 0 .0 4 6 2 .0 3 .0 2 d d L e 2 .8 e r 2 4 r e P e A 1 .5 t t n F C n 2 .6 e U e U c c f - D - o n n N 2 .4 1 .0 a a f i y i t i o d d

s e e y n t 2 .2 i m m e

t 2

2 n 0 .5 n i g g e

t 2 .0 o o l l n i

0 .0 1 .8 C o n tro l C G N C o n tro l C G N

Expression of PPARGC1A (a) and its downstream gene NRF1(b), CPT1B(c), SLC25A19(d), NDUFA4L2 (e), UCP2 (f) in human sample. n= 9 in control, n=6 in CGN. t test.

Supplement Figure 7. Our model of PGC1α-induced podocytes dysfunction and kidney disease

In podocytes, PGC1α increases mitochondrial biogenesis and alters mitochondrial intrinsic properties, including (1) shifts mitochondrial fuel preference, (2) increases mitochondrial fusion and size, and (3) alters electron transport chain subunits assembly, altogether, these changes reprogram cell fate and result in podocyte proliferation and dedifferentiation. Transcriptional factor (TF)

Supplement table 1. List of differentially expressed gene between Control and Nefta-PGC1α mouse glomeruli detected by RNAseq Gene Base Mean log2 Fold Change P value P adj Cox7a1 898.5755 3.824365 9.65E-77 1.49E-72 Atp1a3 1329.264 4.645603 1.44E-71 1.11E-67 Ndrg4 1226.772 4.487786 2.29E-66 1.18E-62 Kirrel2 8140.544 4.41017 8.49E-65 3.28E-61 Mthfd2 509.164 3.1796 1.81E-50 5.61E-47 Hcn2 260.7521 3.499623 3.29E-41 8.49E-38 Cend1 533.3176 3.681548 1.76E-40 3.88E-37 Ppargc1a 3527.44 2.591867 2.68E-40 5.18E-37 Tm6sf1 468.4285 2.645751 2.7E-33 4.64E-30 Idh3a 5283.4 1.990246 6.62E-28 1.02E-24 Gpr162 530.2162 2.348869 1.45E-27 2.04E-24 Got1 4159.253 1.79161 7.79E-26 1E-22 Dsc2 207.8818 2.681008 1.09E-24 1.29E-21 Chchd10 6210.611 1.821575 1.3E-24 1.44E-21 Myl6b 421.6918 2.429305 8.68E-24 8.95E-21 Clec2l 146.679 2.805462 1.31E-22 1.27E-19 Tmprss13 86.96685 3.059017 1.42E-22 1.29E-19 Bcan 110.0747 2.959506 2.69E-22 2.31E-19 Slc41a3 646.787 2.067545 3.06E-22 2.49E-19 Asns 386.892 2.320291 3.21E-21 2.48E-18 Tnfrsf12a 1821.507 2.493598 3.56E-21 2.62E-18 Fndc5 1445.211 1.879443 1.94E-20 1.36E-17 Atf5 1851.33 2.118376 6.5E-20 4.37E-17 Pcolce 10858.27 1.455393 1.73E-19 1.11E-16 Ndufa5 2023.069 1.522956 2.54E-19 1.57E-16 Cs 9324.68 1.417782 2.74E-19 1.63E-16 Tmem35 302.5389 2.217028 4.23E-19 2.43E-16 Trp53inp2 12762.16 1.370049 4.53E-19 2.5E-16 Dlat 4014.676 1.391433 8.03E-19 4.28E-16 St8sia5 66.48737 2.794029 2.05E-18 1.06E-15 Acot2 1381.979 1.980366 2.25E-17 1.12E-14 Atp2b2 320.4693 1.954806 5.94E-17 2.87E-14 Dok7 78.41684 2.58202 6.95E-17 3.26E-14 Got2 2870.829 1.496752 2.1E-16 9.56E-14 Slc25a4 10989.52 1.429829 3.98E-16 1.76E-13 Stbd1 643.3227 1.875897 6.64E-16 2.85E-13 Hspb8 3666.494 1.306192 7.39E-16 3.09E-13 Acot7 1126.256 1.64962 8.68E-16 3.53E-13 Aco2 17638.09 1.291538 1.14E-15 4.51E-13 Cidea 57.78861 2.518784 4.64E-15 1.79E-12 Fuom 670.2155 1.675142 1.16E-14 4.38E-12 Ogdh 19685.37 1.20362 2.34E-14 8.62E-12 8

Mtfp1 508.0732 1.607963 1.41E-13 5.07E-11 Ghitm 13328.52 1.107637 1.47E-13 5.11E-11 Map1b 1331.502 1.541642 1.49E-13 5.11E-11 Pdk4 2956.98 2.291267 1.99E-13 6.69E-11 Tmem38a 474.2643 1.545317 5.18E-13 1.7E-10 Cox5a 3272.596 1.301767 7.71E-13 2.44E-10 Trib3 147.9948 2.192117 7.73E-13 2.44E-10 Kcnn1 170.1885 2.108392 1.44E-12 4.45E-10 Atp5o 3573.51 1.131008 1.81E-12 5.45E-10 H2-Q7 386.9678 -1.75805 1.83E-12 5.45E-10 Chd5 171.4752 2.253789 2.35E-12 6.86E-10 Ndufab1 768.1508 1.288409 4.29E-12 1.23E-09 Eno1 1374.816 1.204752 5.1E-12 1.43E-09 Etfdh 3741.341 1.243482 8.14E-12 2.25E-09 Hpdl 74.56846 2.100107 1.23E-11 3.35E-09 Ndufb6 1180.451 1.18669 1.45E-11 3.88E-09 Hspa9 12340 1.126492 2.42E-11 6.35E-09 P4ha3 62.26041 2.08706 2.89E-11 7.46E-09 Ndufb10 3414.712 1.129774 3.46E-11 8.77E-09 Hk1 5781.684 1.03359 4.19E-11 1.04E-08 Obscn 69.43361 2.09336 5.33E-11 1.31E-08 Ndufs1 5450.06 1.125446 6.21E-11 1.5E-08 Ndufa11 1601.162 1.115552 7.08E-11 1.68E-08 Abca13 1058.007 -1.47272 8.25E-11 1.93E-08 Atp5g1 380.7126 1.458278 1.01E-10 2.34E-08 Spock2 194.1348 1.959635 1.29E-10 2.89E-08 Syt17 620.2823 1.277955 1.29E-10 2.89E-08 Eno1b 543.0175 1.272679 1.58E-10 3.5E-08 Mdh2 9007.365 1.070796 1.62E-10 3.53E-08 Eef1a2 60.26149 2.049154 1.79E-10 3.85E-08 Rin1 341.7687 1.594063 1.95E-10 4.14E-08 Ifrd1 1802.63 1.494539 2.3E-10 4.8E-08 Cox4i1 10536.56 0.984889 3.42E-10 7.05E-08 Adssl1 338.2799 1.660514 3.91E-10 7.96E-08 Atp5g3 11188.23 1.090609 5.23E-10 1.05E-07 Lrfn5 39.22838 1.991232 5.81E-10 1.15E-07 Ndufb9 3989.784 1.099489 7.93E-10 1.55E-07 Ndufs2 7835.564 0.92181 8.92E-10 1.7E-07 Sucla2 4097.963 1.03779 8.89E-10 1.7E-07 Gfm1 2635.209 1.111496 1.16E-09 2.19E-07 Nupr1 4964.527 1.02162 1.19E-09 2.21E-07 Pkm 7269.041 1.00226 1.27E-09 2.34E-07 Slc25a3 16318.56 1.013626 1.31E-09 2.38E-07 Uqcrb 884.1445 1.111799 1.45E-09 2.6E-07 Cox6b1 4532.374 0.980531 1.59E-09 2.83E-07

Kcng2 300.4519 -1.40253 2.11E-09 3.7E-07 Tmem215 40.91442 1.919244 2.16E-09 3.75E-07 Ndufb8 3634.828 1.140643 2.94E-09 5.05E-07 Ndrg2 2669.957 1.430762 3.04E-09 5.17E-07 Ndufv1 5796.075 0.963283 3.37E-09 5.67E-07 Atp5j2 2297.044 1.01251 3.5E-09 5.83E-07 Gars 2581.83 1.087054 3.69E-09 6.01E-07 Uqcrh 3310.721 0.966425 3.67E-09 6.01E-07 Cyc1 8060.04 1.089123 3.75E-09 6.04E-07 Atp5b 55637.13 0.973799 3.88E-09 6.18E-07 Ndufb5 2005.54 0.961791 4.18E-09 6.59E-07 Acadvl 2930.206 1.135081 4.68E-09 7.31E-07 Cox6a1 8067.296 1.024737 4.88E-09 7.55E-07 Ndufs8 2355.48 0.96075 5.79E-09 8.86E-07 Atp5k 1728.819 0.990101 7.06E-09 1.07E-06 Nol3 177.202 1.59667 7.42E-09 1.11E-06 Ndufv2 2953.698 0.95392 1.32E-08 1.96E-06 Ndufaf4 941.6183 1.084771 1.58E-08 2.31E-06 Cdsn 62.43413 1.793137 1.59E-08 2.31E-06 Ndufa10 3534.753 0.898717 2.41E-08 3.49E-06 Cox7b 2782.185 1.025276 3.03E-08 4.33E-06 Atp5d 6608.087 0.890448 3.28E-08 4.65E-06 Hadha 6767.798 1.044238 3.4E-08 4.78E-06 Chchd3 2801.711 0.874925 3.66E-08 5.1E-06 Hapln4 45.2861 1.75986 4.1E-08 5.56E-06 Me3 821.8796 1.273701 4.08E-08 5.56E-06 Osbpl1a 1370.891 1.133136 4.09E-08 5.56E-06 Matn4 119.9404 1.602144 4.98E-08 6.69E-06 Slc26a7 995.2798 -1.35644 6.17E-08 8.23E-06 Cox5b 1315.3 1.049904 6.98E-08 9.23E-06 Ckb 2128.313 1.406125 7.55E-08 9.89E-06 Ndufa4 4732.238 0.916159 8.12E-08 1.05E-05 Nceh1 2893.243 1.024385 8.38E-08 1.08E-05 Plagl1 5049.432 0.931514 9.68E-08 1.24E-05 Pigr 696.6174 -1.11079 9.94E-08 1.25E-05 Mpc2 2628.11 0.958421 1E-07 1.25E-05 Aldh1b1 88.22498 1.605691 9.86E-08 1.25E-05 Uqcrq 2657.443 0.946896 1.05E-07 1.3E-05 Hadhb 742.5289 0.983629 1.12E-07 1.37E-05 Tnfaip2 4019.357 0.915349 1.69E-07 2.06E-05 Sdhb 5355.009 0.884137 1.77E-07 2.14E-05 Hsdl2 2088.304 0.840574 1.89E-07 2.26E-05 Frem1 297.814 1.402588 2.36E-07 2.81E-05 Pdhb 1918.819 0.871733 2.56E-07 3.02E-05 Mrm1 425.2162 1.100581 2.89E-07 3.39E-05

Vegfb 1116.769 0.928172 3.12E-07 3.63E-05 Ndufb7 1861.508 0.880736 3.4E-07 3.92E-05 Slc25a34 526.7743 1.196023 3.72E-07 4.26E-05 Dld 5112.456 0.846189 3.74E-07 4.26E-05 Tfrc 1274.835 1.144224 3.83E-07 4.33E-05 Cox6c 2410.846 0.984686 4.22E-07 4.73E-05 Spata21 34.62854 1.565772 5.08E-07 5.65E-05 Fam134b 3677.389 1.060709 5.21E-07 5.76E-05 Tlr7 451.975 -1.60489 5.3E-07 5.81E-05 Atp5a1 38523.24 0.77136 5.74E-07 6.25E-05 Ppm1l 1404.95 0.954114 5.83E-07 6.31E-05 Trak2 11216.48 0.925545 6.09E-07 6.54E-05 Lypd1 32.4547 -1.5884 6.23E-07 6.64E-05 Hccs 945.6991 0.909511 6.3E-07 6.67E-05 2010107E04Rik 1700.647 0.957715 6.6E-07 6.9E-05 Gm1673 92.82086 1.515902 6.59E-07 6.9E-05 Slc25a5 5054.425 0.804807 6.9E-07 7.07E-05 Pdha1 6924.981 0.816185 6.84E-07 7.07E-05 Comp 57.67562 1.598818 6.87E-07 7.07E-05 Cox10 694.2263 1.122401 7.34E-07 7.47E-05 Grin2d 143.0628 1.301441 8.24E-07 8.33E-05 Ndufc2 1357.61 0.871517 8.68E-07 8.71E-05 Cxcr4 429.9657 -1.16517 8.73E-07 8.71E-05 Immt 5859.86 0.830046 9.47E-07 9.39E-05 Esrra 2654.653 0.798274 9.64E-07 9.49E-05 Mycl 350.7031 1.120316 9.75E-07 9.54E-05 Fh1 3780.02 0.944007 1.06E-06 0.000103 Gdpd1 234.7716 1.239727 1.13E-06 0.000109 Ndufa9 3365.979 0.848303 1.17E-06 0.000112 Pamr1 917.4512 1.436484 1.18E-06 0.000113 Neurl1a 77.70256 1.460664 1.22E-06 0.000115 Coq7 703.8306 0.925098 1.22E-06 0.000115 Sdhd 4480.523 0.829838 1.61E-06 0.00015 Bdh1 1312.265 0.832846 1.71E-06 0.000159 Atp5c1 6711.621 0.770598 1.73E-06 0.00016 Fam129a 576.3212 1.284817 1.76E-06 0.000162 Mpnd 1329.284 0.824244 1.91E-06 0.000175 Btnl9 1132.381 -1.06488 1.96E-06 0.000178 Tmc7 955.0734 -0.93864 2.04E-06 0.000185 Slc7a5 328.465 1.228608 2.22E-06 0.000199 Pdhx 1000.968 0.874377 2.24E-06 0.000201 Zim1 50.58519 1.499728 2.34E-06 0.000207 Stac2 494.3771 1.491523 2.33E-06 0.000207 Gapdh 525.5843 0.947747 2.6E-06 0.000229 Dlst 7999.655 0.81486 2.81E-06 0.000245

Lfng 1660.706 0.850636 2.8E-06 0.000245 Syt15 206.8838 -1.27437 2.93E-06 0.000253 Ddit3 1167.851 1.14832 2.97E-06 0.000255 Hoxd10 244.8141 1.094603 3.14E-06 0.000269 Retsat 2198.347 0.861617 3.32E-06 0.000282 Cox7a2 2982.077 0.83646 3.5E-06 0.000296 Dnajc15 560.3516 0.958747 3.58E-06 0.000301 Oxct1 19301.26 0.972009 3.6E-06 0.000301 Mdh1 15047.01 0.808409 3.63E-06 0.000302 Pfkp 1300.205 1.138444 3.84E-06 0.000318 Uqcr11 1143.177 0.805735 4.03E-06 0.000332 Mrpl34 635.0963 0.890128 4.32E-06 0.000353 Eif4ebp1 509.4691 1.224842 4.35E-06 0.000354 Thsd7a 11626.31 -0.84953 4.54E-06 0.000367 Aqp2 1649.035 -0.77048 4.62E-06 0.000372 Snx31 1222.826 -1.01816 4.77E-06 0.000382 Idh3g 4637.493 0.773542 5.25E-06 0.000417 Myof 8160.714 0.967574 5.25E-06 0.000417 Minos1 2128.391 0.890204 5.46E-06 0.00043 Slc16a1 550.3111 1.24744 5.69E-06 0.000447 Ppfia3 47.12315 1.444035 5.75E-06 0.000449 Uqcr10 2527.062 0.754405 5.81E-06 0.000452 Ndufa8 2169.79 0.828281 5.93E-06 0.000459 Pm20d2 51.99667 1.434584 5.96E-06 0.000459 Ndufb11 2701.199 0.706447 6.42E-06 0.000492 mt-Tp 554.0792 1.018517 6.61E-06 0.000503 Msrb2 515.9675 0.945506 6.77E-06 0.000513 Srxn1 1125.905 0.788296 6.96E-06 0.000525 Uqcrc2 7722.228 0.76318 7.14E-06 0.000536 Ak1 3007.941 0.814229 7.21E-06 0.000538 1700021F05Rik 537.36 0.901387 7.66E-06 0.00057 Gbas 2058.544 0.750526 9.54E-06 0.000706 Slmo2 1159.684 0.770447 9.6E-06 0.000707 Idh3b 5425.934 0.780704 1E-05 0.000734 Mtx2 1408.817 0.743092 1.02E-05 0.000744 Tomm40 1126.754 0.75424 1.12E-05 0.000812 Ptprv 729.3431 1.252106 1.13E-05 0.000817 Ppp1r14b 519.4662 0.92061 1.17E-05 0.000844 Ndufaf5 303.1045 0.960936 1.23E-05 0.000873 Ndufs5 354.4787 0.934342 1.22E-05 0.000873 Fbxo44 379.7743 1.045506 1.27E-05 0.000902 Slc37a2 530.5086 1.1644 1.33E-05 0.000941 Ndufc1 863.5228 0.846308 1.36E-05 0.000953 Pgm1 942.2187 0.834432 1.56E-05 0.00109 Tpi1 6380.348 0.721216 1.59E-05 0.00111

Sars 2771.676 0.698584 1.61E-05 0.001118 Cox6a2 52.72744 1.370327 1.63E-05 0.001127 Ndufa6 2705.556 0.737306 1.72E-05 0.001184 Cycs 172.4328 1.105763 1.76E-05 0.001204 Ndufa1 1524.117 0.76238 1.78E-05 0.001215 Tnfrsf25 136.8876 -1.20824 2.06E-05 0.001396 Tmem246 640.3801 0.807636 2.09E-05 0.001409 Atp5j 3385.526 0.674531 2.21E-05 0.001485 Rab30 136.0493 1.129435 2.3E-05 0.001538 Mrpl47 364.3939 0.977111 2.39E-05 0.001596 Sec16b 241.7169 -1.09339 2.45E-05 0.001628 Rsad1 255.8602 0.983553 2.58E-05 0.001705 Gm266 232.8831 -1.3037 2.86E-05 0.001881 Sfrp1 1970.565 -0.91197 2.94E-05 0.001926 Calr3 286.0431 1.111641 2.96E-05 0.001929 Ndc1 504.7128 0.830176 3.03E-05 0.001969 Prdx3 3067.492 0.742322 3.12E-05 0.002019 Pnpla2 4214.033 0.78491 3.14E-05 0.002021 Tuba1a 1829.634 0.883067 3.17E-05 0.002034 Sod2 4034.155 0.689577 3.46E-05 0.002201 Vdac2 6190.791 0.644949 3.49E-05 0.002201 Sema3g 51714.96 -1.17657 3.48E-05 0.002201 Atp5h 1128.509 0.737803 3.49E-05 0.002201 Ptgds 256.8206 1.308513 3.74E-05 0.002345 2410015M20Ri k 1252.116 0.736597 3.74E-05 0.002345 Ppp3cc 501.7548 0.813993 3.95E-05 0.002462 Lrpprc 2493.019 0.793509 3.97E-05 0.002468 Myo19 649.5969 0.782066 4.05E-05 0.002505 1110001J03Rik 565.8186 0.906468 4.17E-05 0.002568 Lypd6 492.073 -1.07443 4.42E-05 0.002711 Ptprc 244.5057 -1.00482 4.48E-05 0.002741 Pfkm 2015.829 0.709266 4.51E-05 0.002748 Gpr3 34.08355 1.28255 4.6E-05 0.002781 C430049B03Rik 404.0515 0.861733 4.59E-05 0.002781 Sec14l2 148.3904 1.070624 4.97E-05 0.002988 Srrm4 858.5813 1.122537 4.99E-05 0.002988 Cyp2d9 155.6567 -1.21083 5.04E-05 0.003012 Prdx2 2646.806 0.654497 5.41E-05 0.003219 Coq6 555.7104 0.781946 5.8E-05 0.003425 Maff 503.4851 1.239926 5.8E-05 0.003425 Cox8a 6704.124 0.683171 5.87E-05 0.003452 Aars 3374.993 0.676126 5.93E-05 0.003475 Fam162a 998.3415 0.696228 6.35E-05 0.003674 Wnt11 87.18922 1.212849 6.37E-05 0.003674 Pptc7 2070.446 0.665047 6.31E-05 0.003674

Bola3 708.1987 0.804612 6.35E-05 0.003674 Mrpl30 769.5208 0.752144 7.03E-05 0.004042 Pacsin1 27.44319 1.269812 7.15E-05 0.004098 Nars 3772.165 0.613137 7.22E-05 0.004108 Tmem54 1406.86 0.698679 7.21E-05 0.004108 Arhgef19 120.5976 1.165455 7.3E-05 0.004123 Adamts3 58.42151 1.277657 7.29E-05 0.004123 Zfp691 258.9945 0.924462 7.66E-05 0.004308 Sdsl 133.9057 1.093287 8.28E-05 0.004642 Uqcrc1 8491.717 0.741279 8.5E-05 0.004745 Fastkd1 490.8969 0.839928 8.63E-05 0.004782 Itgal 148.1896 -1.06785 8.61E-05 0.004782 Gde1 3514.161 0.677455 8.82E-05 0.004874 Synm 781.1308 -0.71604 8.86E-05 0.004874 Prr15 389.1567 -0.90337 9.38E-05 0.005142 Meis2 4367.466 -0.64534 9.54E-05 0.005213 Ltbp4 3900.514 -0.69605 9.6E-05 0.005227 Uqcrfs1 5062.692 0.711388 0.0001 0.005448 Kcnj1 1574.977 -0.77187 0.000101 0.00546 Myh11 2475.124 -1.00228 0.000103 0.005555 Ndufa12 427.7416 0.813574 0.000104 0.005565 Slc41a2 1413.803 0.781536 0.000104 0.005565 Letm1 3428.991 0.673613 0.000106 0.005644 Tbc1d9 408.7098 1.108138 0.000106 0.005644 Crct1 273.8309 1.123198 0.000107 0.005686 Aldoa 17083.63 0.627953 0.000109 0.005751 Iars 1822.072 0.645804 0.00011 0.005775 Atp1b1 23544.15 0.641301 0.00011 0.005777 Scn4b 512.3969 -0.79243 0.000112 0.005827 Igfbp3 653.9203 -0.72201 0.000117 0.006083 Mrps23 980.0684 0.674462 0.000118 0.006149 Atp5f1 4177.262 0.700802 0.000129 0.006666 Tmx2 577.2646 0.748071 0.000129 0.006666 Atp5e 2753.886 0.634054 0.000132 0.006758 Ppa1 436.7152 0.836234 0.000132 0.006758 Mef2c 1950 -0.67448 0.000133 0.006775 A230052G05Rik 17.54382 1.129453 0.000134 0.006817 Ndufa13 2500.151 0.662316 0.000134 0.006817 Krt7 1828.531 -0.83334 0.000136 0.006874 Dll4 2070.893 -0.75608 0.000139 0.006977 Mfn1 2201.912 0.623145 0.000139 0.006977 RP24-358M18.2 456.5347 -0.90095 0.000142 0.007089 Ptges2 978.2669 0.779062 0.000142 0.007104 Cdkn1a 2639.636 1.122257 0.000147 0.007318 Mrpl15 843.4157 0.722786 0.000154 0.007634

Ccbe1 370.9106 1.173891 0.000155 0.007658 Ndufs6 1020.889 0.734328 0.000156 0.007682 Slc25a39 6857.003 0.627527 0.000158 0.007739 Vwf 89.13652 -1.2031 0.00016 0.007782 Atp6v1b1 689.3563 -0.80889 0.00016 0.007782 Cx3cl1 2720.721 -0.69863 0.000161 0.007782 Sptbn5 670.363 -1.17934 0.00016 0.007782 Ndufs4 1444.833 0.650773 0.00017 0.008213 Them4 348.531 0.794197 0.000171 0.008247 Fbln5 967.1415 -0.67908 0.000176 0.008464 Fam73b 1411.797 0.742022 0.000178 0.008502 Hdac9 265.0137 -0.88024 0.000182 0.008709 Slc13a3 4697.382 -0.68337 0.000187 0.008884 Grsf1 2509.384 0.625457 0.000188 0.008923 Gale 144.6451 0.978448 0.00019 0.008995 Esm1 150.6314 -1.14431 0.000192 0.009042 Mrps18a 913.1917 0.660048 0.000193 0.009057 Mrpl35 656.935 0.691355 0.000201 0.009436 Ushbp1 2425.646 -0.71226 0.000212 0.009911 Mt1 1172.925 0.915118 0.000213 0.009937 Foxc2 2786.154 -0.77509 0.000216 0.010034 Cers6 3599.202 -0.7229 0.000224 0.010353 Cyp26b1 5392.087 1.137148 0.000224 0.010353 Mrpl42 963.2396 0.67638 0.000246 0.011332 S100g 717.3052 -0.7664 0.000251 0.011513 Ighg2b 21.6633 -1.14377 0.000254 0.011635 Timm17a 1340.228 0.677188 0.00026 0.011869 E2f1 869.5626 -0.82258 0.000265 0.012057 Adamts9 1053.369 0.949468 0.000284 0.012886 Afg3l2 2313.942 0.675117 0.000301 0.013617 Slc39a14 773.0135 0.672632 0.000322 0.01452 Masp1 47.25762 1.156834 0.000326 0.014669 Calb1 1370.408 -0.91468 0.000349 0.015661 Rasgrf2 512.2727 -0.74232 0.000352 0.015753 Ung 85.56479 1.131877 0.000354 0.015774 Ptcd2 607.6323 0.697463 0.000356 0.01583 Sdha 13052.73 0.647313 0.000366 0.01619 Sirt3 1146.065 0.666554 0.000366 0.01619 Ackr3 5496.91 0.749534 0.00039 0.017191 Agl 1494.327 0.656157 0.000393 0.01723 Slc19a2 382.0342 0.951095 0.000392 0.01723 Kctd9 758.8676 0.655971 0.000396 0.017303 Tmco4 621.2079 0.755953 0.000404 0.017587 Hspa1b 8142.891 -0.99058 0.000408 0.017733 Pde4b 2059.24 -0.8001 0.000413 0.017909

Fam131c 32.46262 1.130715 0.000424 0.018258 Cars 724.4805 0.698142 0.000424 0.018258 Cluh 4205.019 0.772779 0.000426 0.018321 Endog 332.2461 0.773398 0.000429 0.01839 Timm10 314.3804 0.921904 0.000434 0.01845 Mrpl51 798.4203 0.637858 0.000435 0.01845 Lace1 740.4615 0.694778 0.000435 0.01845 Rpp25 24.20838 1.091346 0.000435 0.01845 Alkbh7 310.9824 0.869631 0.000445 0.018812 Nudt19 2859.905 0.749294 0.00045 0.018979 1110007C09Rik 710.8027 0.764722 0.000456 0.019172 Cldn1 1560.014 1.055258 0.000464 0.019413 Cxcl10 138.1841 1.115848 0.000464 0.019413 Tril 1078.08 -0.6282 0.00047 0.019612 Coq5 960.2572 0.625245 0.000473 0.019677 Clcnkb 1052.287 -0.75449 0.000479 0.019793 Cmc2 380.5115 0.819687 0.00048 0.019793 Tmem106b 6432.846 0.586902 0.000479 0.019793 Phb2 2598.679 0.578033 0.000511 0.021024 Tarsl2 370.098 0.746405 0.000516 0.021166 Ppp1r13l 283.0569 1.02301 0.000519 0.021241 Nfkb2 1536.697 0.661541 0.000523 0.02134 Cldn8 353.6611 -0.82172 0.00053 0.021563 Icam4 27.67144 1.109724 0.000534 0.021668 Coa5 3469.224 0.539491 0.000536 0.02168 Macrod1 437.3865 0.855343 0.000538 0.021706 Filip1 555.7478 -0.82132 0.000545 0.021941 Slc25a22 878.9059 0.617119 0.000549 0.022043 Pi4ka 5344.063 0.61613 0.000575 0.022989 Slit3 609.0933 -0.80256 0.000575 0.022989 Ndufs3 1225.708 0.668244 0.000577 0.02299 Fa2h 1137.87 0.664275 0.000586 0.023288 Ube2m 2180.74 0.562965 0.000592 0.023446 Man2a1 6101.354 0.690023 0.000593 0.023446 Ano8 829.8239 0.671567 0.000601 0.023729 Adamtsl1 174.236 -0.8812 0.000605 0.023812 Foxred1 831.6632 0.620976 0.000628 0.024636 St8sia2 198.5797 1.079568 0.000629 0.024641 Fdx1 896.0442 0.668939 0.000634 0.024769 Magi2 3913.572 -0.9525 0.000638 0.024872 Tiam2 137.138 0.968194 0.000644 0.025037 Hpd 148.3579 -1.07409 0.00065 0.025204 Nrg1 98.94428 1.092563 0.000654 0.025277 Tmem189 2821.58 0.5305 0.000664 0.025626 Gpnmb 80.45 0.964159 0.000671 0.025821

Elovl6 553.3086 0.841789 0.000675 0.025831 Ren1 7120.643 -0.91868 0.000676 0.025831 Gm12892 113.7564 1.084947 0.000675 0.025831 Pla2g4b 23.3102 -1.03274 0.000706 0.026903 Slc38a2 5853.44 0.757589 0.000729 0.027713 mt-Tc 141.6799 1.006832 0.000739 0.028015 Cryab 5692.394 0.767114 0.000747 0.028166 Smpd1 2098.549 0.546334 0.000749 0.028166 Tbcel 2369.831 0.589179 0.000748 0.028166 Pnpt1 780.242 0.618793 0.000754 0.028287 Zfp651 1197.43 0.620096 0.000757 0.028342 Fitm2 775.7939 0.619756 0.000759 0.028342 Samm50 2761.698 0.562372 0.000794 0.029577 Ptcd3 871.2707 0.647119 0.000803 0.029862 Lmo2 1386.071 -0.60076 0.000809 0.030019 Fars2 929.5735 0.617142 0.000826 0.03054 Pik3c2b 490.9971 -0.81477 0.000828 0.03054 Cep68 1129.661 0.640462 0.000829 0.03054 RP23-349J3.8 144.2642 0.890284 0.000833 0.030583 Mrpl19 483.089 0.662278 0.000842 0.030777 Gpx4 1927.497 0.581779 0.000841 0.030777 Arl4c 2034.913 0.867823 0.0009 0.032842 B4galt5 1593.986 0.566209 0.000925 0.033614 Slc25a35 643.2165 0.805807 0.000926 0.033614 Fbxo2 161.3754 1.059855 0.00093 0.033666 Letmd1 1154.067 0.59166 0.000939 0.033927 Dchs1 1726.963 -0.75011 0.000958 0.034448 Mrpl12 1224.742 0.677385 0.000957 0.034448 Adamts8 59.05421 1.049784 0.000963 0.034547 Ndufb2 489.887 0.693859 0.000989 0.035408 Ngb 143.6135 -1.05965 0.000993 0.035469 Cers4 1110.006 -0.74758 0.001 0.035636 St6gal1 2954.279 -0.67801 0.001043 0.037086 1810034E14Rik 90.74155 0.941837 0.001072 0.038011 Ndufb4 154.5115 0.867855 0.001078 0.038101 mt-Tl1 67.10146 1.032961 0.001079 0.038101 Tyro3 1040.913 -0.8961 0.001099 0.03865 Oaf 329.2982 0.713775 0.0011 0.03865 Tufm 265.0978 0.748786 0.001118 0.039197 Hgd 1745.951 -0.6774 0.001151 0.040282 Clic3 4352.275 -0.87959 0.001198 0.041809 Ccr7 28.99809 -1.03358 0.001208 0.042059 Alox12b 34.56413 1.035563 0.001303 0.045269 Crim1 16246.75 0.740612 0.001321 0.04579 Stab1 4331.05 -0.73844 0.001327 0.045917

Cmtm8 708.2951 -0.73538 0.001332 0.045986 Tmem74b 13.96363 0.938233 0.001338 0.046094 Cbx7 1571.197 -0.83333 0.001347 0.046286 Nrtn 110.4804 0.887213 0.001403 0.048121 Atp13a3 7052.835 0.508609 0.001423 0.048674 Oxnad1 674.2947 0.615201 0.001437 0.049046 Mrps24 871.0107 0.581359 0.001442 0.049136 Eif5 5661.997 0.493811 0.001451 0.049332 Abca5 1131.925 -0.59609 0.001456 0.049383 Uqcc2 827.1378 0.598056 0.00146 0.049406 Igdcc4 46.89737 1.01676 0.00147 0.049406 Kctd8 16.33105 0.961861 0.001465 0.049406 Sdpr 7172.211 -0.55428 0.001467 0.049406

Supplement Table 2. qPCR primer sequences Gene Forward Backward Ppargc1a AGTCCCATACACAACCGCAG CCCTTGGGGTCATTTGGTGA Esrra CACTACGGTGTGGCATCCTG ACAGCTGTACTCGATGCTCC Nrf1 CTCGCTGGAAAAAGAAGTG CCGTCCAGGAGTTCAGAGG Nrf2 AAGTGCCCGAAGCCCTCCCT GCCCTTCTGGCGCCGGTTAC Tfam GAGCAGCTAACTCCAAGTCAG GAGCCGAATCATCCTTTGCCT Ndufa10 ACCTTTCACTACCTGCGGATG GTACCCAGGGGCATACTTGC Atp5a1 TCTCCATGCCTCTAACACTCG CCAGGTCAACAGACGTGTCAG Atp5b TCCTGCCAGAGACTATGCG GATGACTGCCACGATTCGC Maoa GCCCAGTATCACAGGCCAC GTCCCACATAAGCTCCACCA Ucp2 CAG CCA GCG CCC AGT ACC CAATGCGGACGGAGGCAAAGC Sod2 GGCCAAGGGAGATGTTACAA GAACCTTGGACTCCCACA Opa1 TGGAAAATGGTTCGAGAGTCAG CATTCCGTCTCTAGGTTAAAGCG Mfn1 ACCTGGAGCAAGAGTTCCAA TGGTGTTTTCAGTGCCAGAG Mfn2 ATGTTACCACGGAGCTGGAC ACTGCTTCTCCGTCTGCAT Drp1 GCGCTGATCCCGCGTCAT CCGCACCCACTGTGTTGA Fis1 TTACCCACTTGCCCCTTTCTT TTGCCCTTCCAGAACTTGGA Slc2a1 ACACTCACCACGCTTTGGTC ACACACCGATGATGAAGCGG Slc2a4 CGTGGCCATCTTCTCTGTGG GCATAGACTCCAAGCCCAGC Hk1 CGGAATGGGGAGCCTTTGG GCCTTCCTTATCCGTTTCAATGG Hk2 TGATCGCCTGCTTATTCACGG AACCGCCTAGAAATCTCCAGA Pkm2 GCCGCCTGGACATTGACTC CCATGAGAGAAATTCAGCCGAG Ldha GCTGCCCCTGCAGAAAATT CACTGGAGTCACAGTTTTCAACGT Pdk4 AAAAAGCGACAACAAACTCCA AACGCAAATCCACCAAGATA Acox1 CTTGGATGGTAGTCCGGAGA TGGCTTCGAGTGAGGAAGTT Acox2 TACCAACGCCTGTTTGAGTG TTTCCAGCTTTGCATCAGTG Acox3 GTGACACGAAGACCCTGCTT TGAACATGGCGAATCCATTA Ldl GGGAGTTTGGCTCCAGAGTTT TGTGTCTTCAGGGGTCCTTAG Acadl TCACCACACAGAATGGGAGA TTTCTCTGCGATGTTGATGC Acadvl GCATCTTGCTCTATGGCACA CACTCGAGGGCTCTGTTAGG Cpt1 CAGGATTTTGCTGTCAACCTC GAGCATCTCCATGGCGTAG Cpt2 CAGCACAGCATCGTACCCA TCCCAATGCCGTTCTCAAAAT Slc1a5 CATCAACGACTCTGTTGTAGACC CGCTGGATACAGGATTGCGG Gls GGGAATTCACTTTTGTCACGA GACTTCACCCTTTGATCACC Glud1 CCCAACTTCTTCAAGATGGTGG AGAGGCTCAACACATGGTTGC Got1 GCGCCTCCATCAGTCTTTG ATTCATCTGTGCGGTACGCTC

Got2 GGACCTCCAGATCCCATCCT GGTTTTCCGTTATCATCCCGGTA c-Myc CAACGACAGCAGCTCGCCCA CCGTGGGGAGGACTCGGAGG Ccnd1 TGGTGGCTGCGATGCA TGCGTTGTGCGGTAGCA Ccne GCGGACACAGCTTCGGGTCT GCGGACTGAAAGGTCGGAGCC Ccna CCCATCGACCGCAGCAAGCA AGGCGGCTCCATGAGGGACA Pcna CACGTATATGCCGAGACCTTAGC CTCCACTTGCAGAAAACTTCACC Wt1 CGGCGGAGTGCTTAGATGAG CTCGCTTCCGTTCCTTCTGT Synpo GGCAGAGGGTGAACGAGTTC GGGCCTCTTGGTTGAGCTTT Nphs1 AGCGTGAGCCCTCACTCGGT CTCCGCCAGAGGAGTCCCCC Nphs2 CACAAGGTTGATCTCCGTCTC GCAGACAGCGTCTATCTCCATT Col1a1 GCCAAGAAGACATCCCTGAA GTTTCCACGTCTCACCATTG Col3a1 ACAGCTGGTGAACCTGGAAG ACCAGGAGATCCATCTCGAC Fn ACAAGGTTCGGGAAGAGGTT CCGTGTAAGGGTCAAAGCAT