Supplementary Information

Supplementary Figure Legends

Figure S1. CARM1 is required for estrogen-induced gene transcriptional activation.

(A) Box plot showing the expression of CARM1 (FPKM) in a cohort of clinical breast cancer

(n=1,102) and normal (n=113) samples from TCGA (The Cancer Genome Atlas).

(B, C) Kaplan Meier survival analyses for DMFS (distal metastasis free survival) (n=1,747) and

OS (overall survival) (n=1,402) of breast cancer patients using CARM1 as input.

(D, E) RNA-seq as described in Figure 1A for specific genes were shown as indicated.

(F) Correlation of CARM1’s effects on whole transcriptome in response to estrogen between two

RNA-seq biological repeats (n=11,816).

(G) Correlation of CARM1 effects on whole transcriptome in response to estrogen based on RNA- seq between two independent siRNAs targeting CARM1 (siCARM1 (1) and siCARM1 (2))

(n=12,234).

(H, I) MCF7 cells as described in (G) were subjected to immunoblotting (IB) (H) and RT-qPCR

(I) analysis to examine the levels of indicated protein and mRNA, respectively. Actin was served as a loading control.

(J) MCF7 cells as described in Figure 1E were subjected to RT-qPCR analysis to examine the expression of genes as indicated.

(K) MCF7 cells as described in Figure 1E were subjected to immunoblotting (IB) analysis to examine the protein levels of CARM1. Actin was served as a loading control.

(L) Genomic DNA was extracted from CARM1 knockout cells, followed by PCR using specific primer sets spanning gRNA targeting region (boxed in blue). The resultant PCR products were subjected to Sanger sequencing as shown. Nucleotide being inserted was highlighted in red. (M) MCF7 cells as described in Figure 1F were subjected to immunoblotting (IB) analysis to examine the protein levels of CARM1 by using anti-specific CARM1 antibodies from two independent vendors as indicated. Actin was served as a loading control.

(N, O) Pol II ChIP-seq signals across specific genes as indicated. Units are mean tags per bin for

16 bins across the transcribed gene region with 2 kb upstream (4 bins of 500 bp each) and 5 kb downstream flanking regions (10 bins of 500 bp each).

(P, Q) Pol II ChIP-seq tag density distribution centered on cognate enhancers of specific genes as indicated (± 4,000 bp).

(R, S) Heat map (R) and box plots (S) representation of the expression levels (FPKM) for genes induced by estrogen and dependent on CARM1 in the cohort of clinical samples, both normal and tumor, as described in (A).

Figure S2. CARM1 is recruited onto ER-bound active enhancers upon estrogen stimulation.

-7 (A) MCF7 cells treated with or without estrogen (E2, 10 M, 1 hr) were subjected to ChIP with anti-CARM1 antibody followed by qPCR analysis with primers specifically targeting enhancer (e) regions as indicated. ChIP signals were presented as percentage of inputs (± s.e.m.).

(B) ChIP-seq tag distribution, including ER, H3K4me1, H3K4me2, H3K4me3, H3K27Ac, P300,

MED1, MED12, H3K9me3 and H3K27me3, centered on all of its own sites (left panels) or estrogen-induced CARM1 sites (right panels) (± 4,000 bp).

(C) Genome browser views of CARM1, ER, H3K4me1, H3K4me2, H3K4me3, H3K27Ac, P300,

MED1, MED12, H3K9me3 and H3K27me3 ChIP-seq in the presence and absence of estrogen on selected active enhancer regions, such as the ones in the vicinity of estrogen-induced coding gene

NRIP1 (left panel on the top), PGR (right panel on the top), SIAH2 (left panel on the bottom) and APOA1 (right panel on the bottom), were shown. Boxed regions indicated active enhancers. ChIP- seq views, except for CARM1, on SIAH2 have been shown in our previous study [1].

Figure S3. Global mapping of CARM1 substrates.

(A) Experimental workflow for global mapping of CARM1 substrates. FASP: filter aided sample preparation.

(B) The distribution of proteins with different number of arginine methylation sites. All the proteins with more than ten methylation sites were shown in oval.

(C) Schematic representation of domain architecture of representative proteins with clustered arginine methylation sites in the “CARM1 methylome”. Arginine methylation sites were shown by matchsticks. CID: CTD-interacting domain; CTD: Spt5 C-terminal domain; G-patch: glycine rich nucleic binding domain; DRBM: double strand RNA-binding domain; PHD: the plant homeodomain zinc finger; TFS2M: transcription factor S-II (TFIIS) domain in the central regions of transcription elongation factor S-II (and elsewhere); RING: ring finger; BBC: B-Box C-terminal domain; BBOX: B-Box-type zinc finger; BROMO: bromo domain; TFS2N: transcription factor

S-II (TFIIS) domain in the N-terminus of transcription elongation factor S-II (and elsewhere);

ZnF_C3H1: (CCCH) type Zinc finger (Znf) domain; KH: K homology RNA-binding domain; IF rod: Intermediate filament (IF) rod domain; WW: domain with 2 conserved Trp (W) residues; FF: a novel motif that often accompanies WW domains and contains two conserved Phe (F) residues.

(D, E) Rates of somatic mutations at (D) and in the vicinity of (E) CARM1 methylation sites in human cancers. Rates of somatic mutations at and in the vicinity of (±5 nucleotides) CARM1- dependent methylation sites (n=589) were significantly higher than those of arginines in the proteome (P = 1.62e-11 and 7.42e-45, respectively).

Figure S4. CARM1 hypermethylates MED12. (A) Human MED12 C-terminus (amino acid (aa) 1616-2177). Arginine residues with mono- methylation only, di-methylation only and both mono- and di-methylation were highlighted in red, green and yellow, respectively.

Figure S5. CARM-mediated methylation is involved in estrogen-induced gene transcriptional activation.

(A) MCF7 cells as described in Figure 5F were subjected to RT-qPCR analysis to examine the expression of CARM1.

(B) MCF7 cells as described in Figure 5F were subjected to immunoblotting analysis to examine the expression of MED12 and CARM1. Actin was served as a loading control.

(C) Correlation of MED12’s effects on transcriptome in response to estrogen based on RNA-seq between two independent siRNAs targeting MED12 (siMED12 (1) and siMED12 (2)).

(D) Correlation of CARM1’s effects on MED12 binding based on ChIP-seq between two independent siRNAs targeting CARM1 (siCARM1 (1) and siCARM1 (2)).

(E, F) MCF7 cells were transfected with control siRNA (siCTL) or two independent siRNAs specific against CARM1 (siCARM1 (1) and siCARM1 (2)) in stripping medium for three days,

-7 and treated with or without estrogen (E2, 10 M, 6 hrs), followed by RT-qPCR (E) and immunoblotting (IB) (F) analysis to examine the mRNA and protein levels of MED12, respectively. Actin was served as a loading control.

(G) MCF7 cells as described in Figure 5J were subjected to RT-qPCR analysis to examine the fold induction by estrogen for selected genes as indicated (± s.e.m., **P<0.01, ***P<0.001).

(H, I) MCF7 cells as described in Figure 5J were subjected to RT-qPCR analysis to examine the expression of CARM1 (H) and MED12 (I) (± s.e.m., ***P<0.001).

Figure S6. TDRD3 is required for estrogen-induced gene transcriptional activation. (A-C) MCF7 cells as described in Figure 6G were subjected to qPCR analysis to examine the expression of selected estrogen-induced genes as indicated.

(D-F) MCF7 cells as described in Figure 6I were subjected to qPCR analysis to examine the expression of selected estrogen-induced genes as indicated.

Figure S7. CARM1 is required for estrogen-induced breast cancer cell growth.

(A) WT and CARM1 KO MCF7 cells were subjected to cell proliferation assay (± s.e.m.,

***P<0.001).

(B) MCF7 cells as described in (A) were subjected to FACS analysis.

(C, D) T47D (C) and BT474 (D) cells transfected with siCTL or siCARM1 were subjected to cell proliferation assay (± s.e.m., *P<0.01, **P<0.01, ***P<0.001).

(E, F) T47D (E) and BT474 (F) cells transfected with siCTL or siCARM1 were subjected to colony formation assay.

Supplementary Table Legend

Table S1. Global mapping of CARM1 substrates. The total number of arginine methylation sites (the sum of mono- and asymmetrical di-methylation sites after removing duplicates), and the number of mono- or asymmetrical di-methyl arginine methylation sites for proteins which had arginine methylation sites identified (sheet 1), had arginine methylation sites on which methylation signals decreased at least two-fold (sheet 2) or completely abolished (sheet 3), and had all its methylation sites abolished (sheet 4) were shown.

Table S2. MS2 spectrum of arginine-methylated peptides in MED12. MS2 spectrum of methylated arginine residues in MED12 C-terminus as described in Figure 4D were shown as indicated. Table S3. Sequence information for all qPCR primers used in the current study. Sequence information of qPCR primers designed to detect the expression of GREB1, NRIP1, PGR, SIAH2,

TSKU, APOA1, ERα, PRSS23, PDCD2L, PSAT1, CAD, CARM1, MED12, TDRD3, SMN, SND1 and SPF30 mRNA, and GREB1, NRIP1, PGR, SIAH2, TSKU and APOA1enhancer RNA (eRNA), and to detect the binding of CARM1 on TFF1 enhancer (e), GREB1 (e), FOXC1 (e), PGR (e),

SIAH2 (e), SMAD7 (e), TSKU (e) and APOA1 (e) regions were shown. F: forward; R: reverse.

Supplementary Reference

1. Gao WW, Xiao RQ, Zhang WJ, Hu YR, Peng BL, Li WJ, et al. JMJD6 Licenses ERalpha-Dependent Enhancer and Coding Gene Activation by Modulating the Recruitment of the CARM1/MED12 Co- activator Complex. Mol Cell. 2018; 70: 340-57 e8.

A 2.5E-46 B C 7 100 100 p=0.0062 p=0.025 6 80 80 5 60 60 4 M (log2) M 40 40

3 OS (%) DMFS (%) DMFS FPK 2 20 CARM1 low expression 20 CARM1 low expression 1 0 CARM1 high expression 0 CARM1 high expression 0 0 20 40 60 80 100 120 0 20 40 60 80 100 120 D F Months G Months 4 4 1kb Pearson Correlation Pearson Correlation chr11:76,493,000-76,511,000 Coefficient = 0.75 Coefficient = 0.75

219.4 ))

3 )) 3 2

siCTL(CTL/E2) 2

219.4 (E siCARM1(CTL/E2) 2 2

TSKU siCTL )/siCTL (E )/siCTL )/ 2 (replicate 2) 2) (replicate E 2

(E 1 1 0.5kb (E

chr11:116,706,000-116,709,000 (siCARM1 change Fold

0 (siCARM1(2) change Fold 0 siCTL(CTL/E2) 0 1 2 3 4 0 1 2 3 4 -12.7 Fold change (siCARM1(1) siCARM1(CTL/E ) Fold change (siCARM1 2 (E )/siCTL (E )) -12.7 (E2)/siCTL (E2)) 2 2 APOA1 (replicate 1) H I J ERα mRNA PRSS23 mRNA PDCD2L mRNA PSAT1 mRNA CAD mRNA siCTL + + - - - - 1.2 3.0 2.5 2.5 3.0 CTL CTL siCARM1(1) - - + + - - 1.0 E 2.0 2.0 E2 - - - - + + 0.8 2 2.0 2.0 siCARM1(2) 1.5 1.5 E - + - + - + 0.6 2 1.0 1.0 0.4 1.0 1.0 IB：CARM1 -60 0.2 0.5 0.5 IB: ERα 0.0 0.0 0.0 0.0 0.0 -60 Normalized Values Normalized Values Normalized Normalized Values Normalized Normalized Values Normalized Normalized Values Normalized IB: Actin -40 K L M WT WT shCTL + + - - 150- KO 150- KO shCARM1 - - + + 100- 100- 70- 70- E - + - + 2 50- 50-

60- CARM1 40- 40- 35- 35- IB: CARM1 KO: CCTCCTCACCATCGGCGACGCCGAACGGCGAGATCCAGCG WT: CCTCCTCACCATCGGCGACGC--GAACGGCGAGATCCAGCG 25- 25- 40- IB: CARM1 (CST) IB: CARM1 (Bethyl) IB: Actin 40- 40- IB: Actin IB: Actin N R S Pol II ChIP-seq tag density P Pol II ChIP-seq tag density 150 siCTL (CTL) 30 siCTL (E ) 2 25 2.42E-05

100 siCARM1 (CTL) 20 Normal Tumor siCARM1 (E ) 6 2 15 50 10 5 Tag density Tag Tag Density Tag 5 4 0 0 4 3 0 5 10 TSKU15 20 25 30 -4000-2000 0 2000 4000 2Kb TSS TTS 5Kb Distance to the center of TSKU enhancer log2) 2 2 Q 1 O Pol II ChIP-seq tag density

Pol II ChIP-seq tag density ( FPKM 150 0 0 siCTL (CTL) 8 siCTL (E ) -1 100 2 6 siCARM1 (CTL) -2 siCARM1 (E2) 4 50 g density 2 Ta Tag Density Tag Genes i) induced by E ; ii) 0 2 0 dependent on CARM1 GREB1 -4000-2000 0 20004000 (n=461) 2Kb TSS TTS 5Kb Distance to the center of GREB1 enhancer Figure S1 A B Centered on all of its Centered on E2-induced own sites CARM1 sites 0.7 CTL 40 CTL 40 E2 0.6 E2 ERα ERα 20 20 0.5 0.4 0 0 -4000 0 4000 -4000 0 4000 0.3 4.0 4.0

% of Inputs of % 0.2 H3K4me1 H3K4me1 0.1 2.0 2.0

0 0.0 0.0 -4000 0 4000 -4000 0 4000 4.0 4.0 H3K4me2 H3K4me2 2.0 2.0 C 40kb chr21:16,313,722-16,625,276 100kb chr11:100,798,522-101,403,488 0.0 0.0 12 CARM1/CTL 7.92 -4000 0 4000 -4000 0 4000 12 CARM1/E 7.92 82 2 5.0 5.0 ERα/CTL 52 82 ERα/E 52 H3K4me3 H3K4me3 3.5 2 H3K4me1/CTL 3.43 2.5 2.5 3.5 H3K4me1/E 3.43 9.96 2 H3K4me2/CTL 9.31 9.96 0.0 0.0 H3K4me2/E 9.31 17 2 -4000 0 4000 H3K4me3/CTL 19 -4000 0 4000 17 H3K4me3/E 19 14 14 37 2 H3K27Ac/CTL 16 37 H3K27Ac/E 16 H3K27ac H3K27ac 16 2 7 P300/CTL 13 7 16 P300/E 13 7.87 2 MED1/CTL 6.98 0 7.87 0 MED1/E 6.98 55 2 -4000 0 4000 -4000 0 4000 MED12/CTL 27 55 MED12/E 27 14 14 30 2 H3K9me3/CTL 30 P300 P300 30 H3K9me3/E 30 2 7 7 30 H3K27me3/CTL 30 30 30 H3K27me3/E2 0 0 NRIP1 PGR -4000 0 4000 -4000 0 4000 5 5 MED1 MED1 5kb chr3:150,448,455-150,488,027 1kb chr11:116,697,018-116,713,847 2.5 2.5 7 20 CARM1/CTL 7 20 CARM1/E 0 0 2 79 92 ERα/CTL -4000 0 4000 79 -4000 0 4000 92 ERα/E 2 4 24 24 9 H3K4me1/CTL 4 9 H3K4me1/E MED12 2 8 MED12 16 H3K4me2/CTL 8 12 12 16 H3K4me2/E 2 4 39 H3K4me3/CTL 4 39 H3K4me3/E 0 0 2 16 106 H3K27Ac/CTL -4000 0 4000 -4000 0 4000 16 106 H3K27Ac/E 2 19 3.0 3.0 36 P300/CTL 19 36 P300/E 2.0 H3K9me3 2.0 H3K9me3 2 8 10 MED1/CTL 8 10 MED1/E 1.0 1.0 2 41 150 MED12/CTL 41 0.0 0.0 150 MED12/E2 30 H3K9me3/CTL 30 -4000 0 4000 -4000 0 4000 30 30 H3K9me3/E2 3.0 3.0 30 H3K27me3/CTL 30 30 H3K27me3/E 30 2.0 H3K27me3 H3K27me3 2 1.5 1.0 SIAH2 APOA1 0.0 0.0 -4000 0 4000 -4000 0 4000

Figure S2 A High pH reversed-phase Wild type (WT) fractionation (Arg0, Lys0) Protein extraction FASP Lys-C/Trypsin digestion 600

me 500 R me me me 400 R R R R R 300 me me Mix 1:1 me mAU 200 R 100 me R R R me R me 0 CARM1 Knockout (KO) R me me 10 20 30 40 50 60 70 (Arg6, Lys6) min

Proteome Intensity m/z (20 fractions) Light Heavy Methylation antibody enrichment LC-MS/MS

me me me me R R R R R R me me me me R R R R R R me me me me Methylome (10 fractions) C 1 CID

PCF11 17 139 1555 B 1104,1093,1086,1074,1065,1062,1051,877,864,833,820 Proteins with arginine methylation sites (n=780) XRN2 1 950 922,921,895,883,880,870,864,847,824 450 420 HNRNPK CTD 400 SRRM2 XRN2 DIDO1 1 563 682 2752 350 KRT8 TRIP6 2392,2384,2370,2342,2288,2274,2246,2231,2221 PCF11 23712426 300 RBMX WDR33 SON 250 RBMXL1 1 23052351 180 YLPM1 KRT18 G-patch DRBM of proteins 200 SRRM2 HNRNPA1 TAF15 1031,1022,1015,996,989,950,943 150 CHTF8 PHD TFS2M DIDO1 100 68 1 270 320 672 773 2240

Number 39 1993,1982,1845,1835,1824,1775 50 14 16 7 12 9 2 4 2 2 3 1 1 0 MED12 L LS PQL Opa 1 2 3 4 5 6 7 8 9 10 11 12 13 15 16 20 1 500 1650 2050 2177 Number of arginine methylation sites per protein 1912,1910,1899,1862,1859,1854 1 1127 TRIM33 PHD (124-185; 889-932) BBC (319-445) RING (217-279) BBOX (271-312) BROMO (959-1082) 591,577,568,555,515 TFS2N ZnF PPP1R10 940 D E 1 74 146 907933 729,726,723,688,665 Any R in human proteome Any AA in vicinity of R in human proteome CARM1-modified R AA in vicinity of CARM1-modified R FAM120A 1 1118 30 27.15 12 10.34 984,982,908,886,884 KH 25 10 KHDRBS1 1 154 252 443 20 17.07 8 340,331,325,291,289 6.02 quency (%) quency 15 6 C6orf132 1 1188 10 4 1127,1119,1053,1044,1038

KRT19 IF rod 5 2 1 80 391 400 Mutation fre Mutation Mutation frequency (%) frequency Mutation 90,51,43,32,24 0 0 R AA surrounding R 1 957 (±5 AA) PRPF40A WW (141-173; 182-214) FF (393-447; 460-519; 527-587; 607-667; 743-799) 52,51,31,9,2 Figure S3 A MED12 (NP_005111.2) aa1616-2177 LAKKLQKELGERQSDSLEKVRQLLPLPKQTRDVITCEPQGSLIDTKGN KIAGFDSIFKKEGLQVSTKQKISPWDLFEGLKPSAPLSWGWFGTVRVD RRVARGEEQQRLLLYHTHLRPRPRAYYLEPLPLPPEDEEPPAPTLLEP EKKAPEPPKTDKPGAAPPSTEERKKKSTKGKKRSQPATKTEDYGMG PGRSGPYGVTVPPDLLHHPNPGSITHLNYRQGSIGLYTQNQPLPAGGP RVDPYRPVRLPMQKLPTRPTYPGVLPTTMTGVMGLEPSSYKTSVYRQ QQPAVPQGQRLRQQLQQSQGMLGQSSVHQMTPSSSYGLQTSQGYT PYVSHVGLQQHTGPAGTMVPPSYSSQPYQSTHPSTNPTLVDPTRHLQ QRPSGYVHQQAPTYGHGLTSTQRFSHQTLQQTPMISTMTPMSAQGVQ AGVRSTAILPEQQQQQQQQQQQQQQQQQQQQQQQQQQYHIRQQQ QQQILRQQQQQQQQQQQQQQQQQQQQQQQQQQHQQQQQQQAAP PQPQPQSQPQFQRQGLQQTQQQQQTAALVRQLQQQLSNTQPQPST NIFGRY

Figure S4 A B C )) 4 Pearson Correlation 2

(E Coefficient = 0.76

3 CARM1 mRNA + + - - siCTL

siCTL )/

1.2 2 1.0 siMED12 - - + +

0.8 E2 - + - + 2 0.6 CTL 180- 0.4 E2 IB: MED12 0.2 60- 1 0.0 IB: CARM1

Normalized Values Normalized 40- IB: Actin 0 0 1 2 3 4 Fold change (siMED12(2) (E (siMED12(2) change Fold Fold change (siMED12(1) (E2)/siCTL (E2)) D E 3 Pearson Correlation Coefficient = 0.54 MED12 mRNA

(siCARM1(2) (siCARM1(2) 1.4 2 )) 1.2 2 1.0 CTL 0.8 E2 0.6 zed Values zed tag density tag

)/siCTL (E )/siCTL 1 0.4 2 0.2 (E seq - 0.0 Normali ChIP 0 0 1 2 3

MED12 MED12 ChIP-seq tag density (siCARM1(1) (E2)/siCTL (E2))

F G TSKU mRNA APOA1 mRNA 8 3.5 siCTL + + - - - - *** *** ** ** ** 7 3 siCARM1(1) - - + + - - 6 6.62 - - - - + + 2.5 2.7 siCARM1(2) 5 2 2.32 /CTL) - + - + - + /CTL) 2 E2 2 4 *** 1.5 3 1.51 180- 3.43 FC (E FC FC (E FC 2 1 1.19 IB: MED12 2.32 2.34 0.5 40- 1 IB: Actin 0 0 shCTL + Vector shMED12 + MED12 (WT) shMED12 + Vector shMED12 + MED12 (R1899A) H I CARM1 mRNA MED12 mRNA 1.4 CTL 120 CTL 97.52

1.2 E2 100 E2 84.44 1 80 0.8 60 52.07 0.6 43.63 40 0.4 *** 20 1.45 0.35 Normalized Values Normalized Normalized Values Normalized 0.2 1 0.42 0 0 shCTL + + ------shCTL + + ------shMED12 - - + + + + + + shMED12 - - + + + + + + Vector + + + + - - + + Vector + + + + - - + + MED12 (WT) - - - - + + - - MED12 (WT) - - - - + + - - MED12 (R1899A) ------+ + MED12 (R1899A) ------+ + Figure S5 A GREB1 mRNA NRIP1 mRNA PGR mRNA SIAH2 mRNA TSKU mRNA APOA1 mRNA SMN mRNA 20.0 5.0 100.0 7.0 3.0 8.0 1.4 * CTL 6.0 1.2 4.0 80.0 2.5 E lues

15.0 lues 6.0 2 5.0 2.0 1.0 3.0 60.0 4.0 0.8 10.0 1.5 4.0 2.0 3.0 0.6 40.0 1.0 5.0 2.0 2.0 0.4 1.0 20.0 1.0 0.5 0.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Normalized Values Normalized Normalized Values Normalized Va Normalized Values Normalized Values Normalized Values Normalized Normalized Va Normalized

B GREB1 mRNA NRIP1 mRNA PGR mRNA SIAH2 mRNA TSKU mRNA APOA1 mRNA SND1 mRNA 15.0 6.0 60.0 7.0 4.0 6.0 1.4 * * * CTL 5.0 50.0 6.0 5.0 1.2 3.0 E2 10.0 4.0 40.0 5.0 4.0 1.0 4.0 3.0 30.0 2.0 3.0 0.8 3.0 5.0 2.0 20.0 2.0 0.6 2.0 1.0 0.4 1.0 10.0 1.0 1.0 0.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Normalized Values Normalized Normalized Values Normalized Values Normalized Values Normalized Values Normalized Values Normalized Values Normalized

C GREB1 mRNA NRIP1 mRNA PGR mRNA SIAH2 mRNA TSKU mRNA APOA1 mRNA SPF30 mRNA 15.0 5.0 40.0 7.0 3.5 6.0 1.5 CTL * * * 6.0 3.0 5.0 E 4.0 30.0 ues ues ues ues 2 10.0 5.0 2.5 4.0 1.0 3.0 4.0 2.0 20.0 3.0 2.0 3.0 1.5 5.0 2.0 0.5 10.0 2.0 1.0 1.0 1.0 0.5 1.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Normalized Values Normalized Values Normalized Values Normalized Val Normalized Val Normalized Val Normalized Val Normalized

D GREB1 mRNA NRIP1 mRNA PGR mRNA SIAH2 mRNA TSKU mRNA APOA1 mRNA SMN mRNA 8.0 5.0 20.0 5.0 8.0 2.5 80.0 * * * CTL 4.0 4.0 2.0 6.0 15.0 6.0 60.0 E2 3.0 3.0 1.5 4.0 10.0 4.0 40.0 2.0 2.0 1.0 2.0 1.0 5.0 1.0 2.0 0.5 20.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Normalized Values Normalized Values Normalized Values Normalized Values Normalized Values Normalized Values Normalized Values Normalized E GREB1 mRNA NRIP1 mRNA PGR mRNA SIAH2 mRNA TSKU mRNA APOA1 mRNA SND1 mRNA 8.0 5.0 25.0 6.0 7.0 2.5 20.0 * * * CTL 6.0 4.0 20.0 5.0 2.0 E 6.0 5.0 15.0 2 alues alues 4.0 3.0 15.0 4.0 1.5 4.0 3.0 10.0 2.0 10.0 3.0 1.0 2.0 2.0 2.0 5.0 1.0 5.0 1.0 1.0 0.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Normalized Values Normalized V Normalized V Normalized Values Normalized Values Normalized Values Normalized Normalized Values Normalized

F GREB1 mRNA NRIP1 mRNA PGR mRNA SIAH2 mRNA TSKU mRNA APOA1 mRNA SPF30 mRNA 10.0 5.0 35.0 6.0 7.0 2.5 200.0 * * * CTL 30.0 5.0 6.0 8.0 4.0 2.0 150.0 E2 25.0 4.0 5.0 6.0 3.0 20.0 4.0 1.5 3.0 100.0 2.0 15.0 3.0 1.0 4.0 2.0 10.0 2.0 50.0 2.0 1.0 5.0 1.0 1.0 0.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Normalized Values Normalized Normalized Values Normalized Values Normalized Values Normalized Values Normalized Values Normalized Normalized Values Normalized

Figure S6 A B 120

2.0 WT 100 1.8 CARM1 (KO) 18.23 15.37 *** 1.6 80 1.4 *** 28.55 G2-M 34.88 S 1.2 ** 60

1.0 * G0-G1 % of Cells of % 0.8 40 0.6 56.08 20 46.89 0.4 Normalized Values Normalized (Absorbance nm) 490 (Absorbance 0.2 0 0.0 WT + - Day: 0 1 2 3 CARM1 (KO) - +

C D T47D BT474 0.4 0.3 siCTL siCTL siCARM1 siCARM1 0.35 0.28 *** ** 0.3

0.26 ** * 0.25 0.24 0.2

Normalized Values Normalized 0.22 Normalized Values Normalized

0.15 nm) 490 (Absorbance (Absorbance nm) 490 (Absorbance

0.1 0.2 Day: 1 2 3 4 Day: 1 2 3 4

E F shCTL shCARM1 shCTL shCARM1

T47D BT474

Figure S7