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Supplemental Figure 1 Functional Proteomic Profiling of Cells Treated with 2DG, AICAR, Metformin, Alone Or in Combination

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Supplemental Figure 1 Functional Proteomic Profiling of Cells Treated with 2DG, AICAR, Metformin, Alone Or in Combination Supplemental Figure 1 Functional proteomic profiling of cells treated with 2DG, AICAR, metformin, alone or in combination. The heatmap represents the normalized intensities of the RPPA experiments for 139 markers. 1 Supplemental Figure 2 Supplemental Figure 5. Cell permeable exogenous energy substrate methyl pyruvate (MP) rescues cell death with 2DG and metformin treatment. (A) Representative photomicrographs of GFP-LC3 U2OS cells 60 hours after treatment of indicated agents. Image captured by high-content image analysis InCell® through brightfield and GFP channels (left 3 columns and right column, respectively).(B) Automated quantitation of autophagosome formation by InCell® system. Data presented mean ± S.D. (n=3). (C) Representative photomicrographs. (D) Trypan blue exclusion assay. Cells were incubated with MP in combination of 2DG and metformin. Data presented mean ± S.D. (n=3) and representative of at least three independent experiments. 2 Supplemental Figure 3 AMPK null MEF cells are more sensitive to glucose deprivation. (A) Cell death in MEF cells lacking functional AMPK catalytic subunit (AMPKα-/-) with glucose deprivation. Representative photomicrographs. Scale bar, 100µm. (B) Cell death in MEF cells lacking functional AMPK catalytic subunit (AMPKα-/-) with 2DG treatment. Data presented mean ± S.D. (n=3) and representative of at least three independent experiments. Representative photomicrographs. Scale bar, 50µm. 3 Supplemental Table 1 The expression of genes in the oxidative phosphorylation of GO category. ANOVA#1 is 6 group one-way ANOVA analysis of gene expression profiles between control, DG, MET, AICAR, DG+MET and DG+AICAR treated groups. ANOVA#2 is 4 group one-way ANOVA analysis of gene expression profiles between control, DG, MET and DG+MET treated groups. ANOVA#3 is 4 group one-way ANOVA analysis of gene expression profiles between control, DG, AICAR and DG+AICAR treated groups. Unique id Gene ANOVA#1 ANOVA#2 ANOVA#3 CONTROL DG MET DG+MET AICAR DG+AICAR Symbol p-value p-value p-value Average Average Average Average Average Average ILMN_4640 ATP5A1 0.0188541 0.1098613 0.2614859 8054.54 8582.96 7158.07 7595.93 9026.75 9165.37 ILMN_759 ATP5B 0.4165370 0.4981272 0.6091699 25599.25 26988.30 24488.94 26314.57 27863.65 27228.22 ILMN_13957 ATP5C1 0.0232545 0.6381015 0.0400105 9385.14 9032.39 8950.07 9416.07 10252.74 10490.25 ILMN_6051 ATP5D 0.0001078 0.4118189 0.0000163 648.46 599.57 685.19 622.92 799.73 860.71 ILMN_6639 ATP5D 0.0008844 0.4702401 0.0015769 1735.99 2077.18 1965.55 1876.10 2671.42 2766.13 ILMN_30107 ATP5E 0.0156013 0.5385018 0.1015114 23953.48 24161.64 21954.74 22642.44 27923.37 28112.51 ILMN_138139 ATP5F1 0.0015518 0.3279987 0.0007444 3081.53 3059.29 2981.43 3647.11 4329.79 4307.38 ILMN_1123 ATP5G1 0.0000001 0.0091483 0.0000189 4676.14 4063.85 4428.20 3541.58 5871.79 6071.02 ILMN_1759 ATP5G1 0.0651569 0.5338593 0.0447280 446.32 454.80 433.43 562.28 648.17 644.39 ILMN_137008 ATP5G2 0.0112787 0.2057288 0.0056689 1321.11 1406.66 1407.21 1500.62 1578.80 1617.74 ILMN_137251 ATP5G3 0.0105917 0.2858730 0.0786700 179.13 189.42 168.62 169.78 209.33 206.30 ILMN_26187 ATP5H 0.0000164 0.4868743 0.0004874 9739.41 9735.32 9481.48 9190.15 11749.84 11707.60 ILMN_29612 ATP5H 0.0020849 0.4487966 0.0109559 16315.55 16102.16 16213.79 15147.28 18569.72 17796.99 ILMN_14284 ATP5I 0.0029753 0.2844527 0.0933737 3073.11 2924.30 2702.98 2752.57 3338.22 3430.78 ILMN_28836 ATP5J 0.0003676 0.7278698 0.0040797 3763.41 3828.60 3356.93 3596.30 5289.28 5601.05 ILMN_29169 ATP5J 0.0000041 0.6911361 0.0000337 1058.39 1209.86 1070.15 1144.14 2014.41 2012.60 ILMN_16064 ATP5L 0.0000423 0.6979367 0.0002203 8298.49 7499.23 7750.90 7827.29 10482.44 10947.07 ILMN_10205 ATP5O 0.2264226 0.1019279 0.3830646 22208.26 20658.48 21312.01 19618.71 20534.30 20709.20 ILMN_139158 ATP6AP1 0.0561817 0.0443753 0.0646397 5561.01 4912.78 4717.44 4330.67 4586.90 4878.54 ILMN_138088 ATP6V0A1 0.0005885 0.0005109 0.0460412 3364.20 3278.28 2727.98 2146.07 2733.64 2846.34 ILMN_23163 ATP6V0A2 0.1814205 0.0509247 0.8914382 564.85 592.77 515.48 612.46 586.04 592.60 ILMN_137821 ATP6V0A4 0.8807447 0.9910077 0.6919854 89.94 90.00 91.23 90.94 95.34 90.01 ILMN_137438 ATP6V0C 0.0017489 0.0075513 0.9451226 15422.64 15000.17 13609.27 12355.45 15344.12 15336.75 ILMN_512 ATP6V0C 0.0000555 0.6516406 0.0000776 477.64 511.75 509.67 524.36 652.07 706.78 ILMN_139373 ATP6V0D1 0.0504860 0.0461080 0.6954964 2802.22 2984.32 2297.70 2588.30 2717.50 2929.37 ILMN_19631 ATP6V0D2 0.2399318 0.2288159 0.6552156 114.23 115.63 118.69 124.20 110.25 117.58 ILMN_30226 ATP6V1A 0.4676691 0.7759381 0.5028699 3454.89 3705.18 3424.40 3325.51 3934.00 3776.02 ILMN_138896 ATP6V1B1 0.3121110 0.2358870 0.4112678 91.81 96.87 86.68 91.22 90.22 90.94 ILMN_137342 ATP6V1B2 0.0020881 0.4550925 0.0001087 3382.33 3232.00 3261.36 3086.33 2902.03 2722.04 ILMN_12844 ATP6V1C1 0.2596007 0.1720159 0.3652393 396.33 464.40 376.05 421.65 443.12 428.12 ILMN_139249 ATP6V1C2 0.9248466 0.8177115 0.9335827 92.07 89.34 87.95 89.33 90.65 91.55 ILMN_26737 ATP6V1D 0.0040097 0.0019098 0.1476213 4200.70 3963.70 4970.09 4845.44 4406.68 4485.98 ILMN_138938 ATP6V1E1 0.0035937 0.0165064 0.0135534 5854.23 5390.25 5407.73 4538.73 4623.25 4705.80 ILMN_5551 ATP6V1E2 < 1e-07 0.0000006 0.0000001 324.62 418.50 211.69 216.77 209.33 192.84 ILMN_20954 ATP6V1F 0.6582436 0.5656186 0.6378461 922.58 963.97 986.73 890.91 954.08 881.76 ILMN_18395 ATP6V1H 0.1231150 0.9708487 0.0912476 2469.90 2509.88 2427.31 2519.28 2130.12 2085.60 ILMN_8500 ATP7A 0.0036093 0.0027375 0.1920515 195.40 206.63 163.13 160.50 188.47 176.47 ILMN_693 C16orf7 < 1e-07 0.0000903 < 1e-07 796.70 797.79 565.30 494.22 303.93 303.83 4 ILMN_693 C16orf7 < 1e-07 0.0000903 < 1e-07 796.70 797.79 565.30 494.22 303.93 303.83 ILMN_21275 COX10 0.0086005 0.1158488 0.0016722 292.03 315.12 359.24 359.79 375.01 406.82 ILMN_18159 DLD 0.3214587 0.2241227 0.7328064 1577.10 1714.22 1563.42 1861.68 1590.50 1622.96 ILMN_15537 LOC92270 < 1e-07 0.5223037 < 1e-07 249.52 263.88 245.57 243.23 163.58 153.64 ILMN_9008 MSH2 0.0862507 0.0962614 0.0503620 654.36 689.45 714.02 837.89 796.82 802.92 ILMN_24450 NDUFA1 0.0060415 0.5262088 0.0406215 9761.53 10035.24 8906.94 9076.52 11254.42 12227.68 ILMN_7463 NDUFA10 0.0526025 0.6650664 0.0600442 2010.85 2036.59 2070.85 1884.52 2392.49 2372.27 ILMN_5263 NDUFA2 0.0068109 0.6209785 0.0117761 2911.20 2661.95 2754.55 2641.70 3434.37 3289.50 ILMN_6292 NDUFA3 0.0294236 0.8951572 0.0388198 3630.94 3257.08 3319.69 3169.52 4940.93 4861.77 ILMN_3753 NDUFA4 0.0492255 0.0521455 0.1602943 22596.25 21910.18 20965.69 18922.09 22059.20 19571.48 ILMN_9798 NDUFA5 0.1559223 0.6494420 0.0874642 464.71 526.18 532.30 510.44 597.87 638.38 ILMN_29741 NDUFA6 0.0000082 0.0134952 0.0003562 1040.60 886.53 944.81 830.84 1203.06 1023.36 ILMN_3334 NDUFA7 0.0349294 0.0318012 0.6089783 1755.15 1687.76 1660.57 1342.42 1653.52 1587.66 ILMN_23439 NDUFA8 0.0314655 0.0587495 0.5905542 4856.80 4659.26 4185.15 3977.75 4974.15 4572.75 ILMN_6998 NDUFA9 0.0000360 0.9516733 0.0000726 3404.00 3451.66 3498.08 3523.88 4530.29 4510.40 ILMN_14128 NDUFAB1 0.0220803 0.3569305 0.0717519 12753.94 12098.68 11954.75 11145.36 14133.56 13522.36 ILMN_13182 NDUFAF1 0.0000001 0.0001336 0.0000004 483.79 419.19 381.59 339.25 314.49 298.81 ILMN_28096 NDUFB1 0.0004769 0.7871930 0.0016686 172.30 177.71 168.65 166.39 204.05 219.48 ILMN_6518 NDUFB10 0.0000004 0.0000657 0.5894113 8072.25 8152.18 7228.18 6126.65 8490.79 8390.27 ILMN_20348 NDUFB2 < 1e-07 0.0000166 0.0121172 9343.07 8707.80 7494.43 6568.15 10530.30 10351.33 ILMN_22320 NDUFB3 0.0018051 0.7941043 0.0006057 1708.75 1598.12 1708.69 1845.25 2507.87 2512.94 ILMN_9342 NDUFB6 0.0000095 0.4416394 0.0003502 1826.33 1799.51 1876.63 1677.09 2368.97 2429.21 ILMN_6301 NDUFB7 0.0000050 0.0409234 0.0063004 4711.21 4664.12 3938.97 3841.51 5877.11 5978.05 ILMN_27115 NDUFB8 0.0010014 0.0168842 0.1998357 10544.66 9831.38 9599.49 8661.34 10728.02 10753.05 ILMN_3456 NDUFB9 0.5921585 0.7728421 0.5937660 10445.10 9814.80 9843.53 9885.22 10749.28 10707.65 ILMN_27316 NDUFC1 0.0006643 0.5122531 0.0015198 1164.21 1252.78 1224.43 1136.06 1540.71 1477.56 ILMN_12753 NDUFC2 0.0039581 0.0063616 0.9200482 182.44 191.80 148.85 137.64 193.38 191.48 ILMN_19530 NDUFS1 0.0052885 0.0172555 0.8006122 353.20 364.58 294.88 314.10 370.35 379.68 ILMN_9109 NDUFS2 0.6644936 0.6546265 0.3175771 763.33 629.68 676.78 689.80 625.04 615.33 ILMN_6465 NDUFS3 0.0312775 0.8891099 0.0293895 5618.98 5553.10 5519.77 5776.57 6482.02 6546.47 ILMN_1167 NDUFS4 0.1779066 0.0849078 0.3837569 5959.26 6489.39 5733.53 6379.37 6097.38 6371.71 ILMN_6762 NDUFS5 0.0109388 0.5557584 0.0130726 5233.62 6580.92 5712.98 5488.67 8540.73 8264.10 ILMN_138215 NDUFS6 0.0002488 0.0112062 0.0983676 12910.16 13245.16 10724.86 10593.81 15180.90 15173.44 ILMN_25892 NDUFS7 0.3064088 0.5962298 0.4744599 2802.68 2752.11 2605.09 2460.22 2842.34 3081.71 ILMN_24953 NDUFS8 0.0006076 0.0094753 0.9077275 10976.78 11057.73 9738.85 9484.97 11283.91 11263.19 ILMN_8645 NDUFV1 0.9141283 0.9546225 0.7887321 2831.48 2703.02 2784.22 2695.77 3002.55 2939.02 ILMN_5793 NDUFV2 0.0312722 0.1745021 0.2819397 2647.20 2775.57 2683.83 2366.25 3032.91 2789.78 ILMN_17835 NDUFV3 0.0416345 0.0219817 0.0308921 210.70 184.05 169.67 175.09 175.02 176.99 ILMN_1502 TAZ 0.0022675 0.1810433 0.0005116 255.53 228.71 228.51 212.81 187.18 190.06 ILMN_29643 TAZ 0.1163138 0.1437879 0.0592035 185.15 158.23 190.69 168.86 183.86 188.68 ILMN_1306 TCIRG1 0.0052327 0.1934532 0.0127967 284.46 269.95 292.25 268.15 296.54 315.33 ILMN_3323 UCRC < 1e-07 0.3344934 0.0000004 914.78 816.55 874.84 803.13 1435.48 1487.42 ILMN_10392 UQCR 0.1707982 0.0341479 0.1363116 197.04 157.47 180.80 167.50 169.44 181.57 ILMN_26282 UQCRB 0.0076585 0.6253715 0.0130910 2148.64 2524.76 2062.12 2322.73 3269.57 3377.66 ILMN_20071 UQCRC1 0.4487900 0.6439481 0.2671411 2970.41 2623.89 2862.25 2666.24 3232.80 3091.10 ILMN_138507 UQCRH 0.0652144 0.6690911 0.0735542 9867.47 9832.76 10239.69 9216.21 11532.36 11682.05 5 Supplemental Table 2 The expression of genes in the TCA of KEGG category.
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