H3r17me2a

TC100 insect medium (Ref: IML007), PBS (Ref: TL1006), Trypsin-EDTA solution 10× (Ref: TCL070) and Antibiotic antimyotic solution 100× (Ref: A002A) were obtained from HIMEDIA. The lipofectamine used for transfection was obtained from Invitrogen (Ref: 11668019). DMEM powder (Ref: D1152), MTT (Ref: M5655) and SAM (Ref: A7007) were obtained from Sigma-Aldrich (St. Louis, Missouri, USA). TRIzol reagent (Ref: 15596018) was obtained from ambion life technologies. The TRIPZ-shRNA plasmids used for stable cell line generation were obtained from Dharmacon (Ref: CARM1 sh -V3THS_319980 and YY1 sh-V2THS_219592). The commercial antibodies used in the study include Tubulin from calbiochem (Ref: CP06), CARM1 from abcam (Ref: Ab84370), YY1 from abcam (Ref: Ab12132) and H3R17me2a from Millipore (07–214). Tritiated SAM was obtained from Perkin-Elmer (Ref: NET155250UC). Luciferase assay buffer (Ref: E151A) and reporter lysis buffer (Ref: E397A) were purchased from Promega (Madison, Wisconsin, USA). CARM1 inhibitor (PRMT inhibitor V) was purchased from Calbiochem-Millipore (Ref: 217531).

D4H5, D6A8 and D10F7 were generated against degenerate peptides containing four asymmetric dimethylarginine (ADMA) residues XXXXRme2aXXRme2aXXXXRme2aXXX Rme2aX by Cell Signaling Technology (CST). D4H5 and D6A8 have been previously described20 (link) and are commercially available as part of an ADMA antibody mixture (CST, Catalog #13522). Anti CAS3 antibodies were custom raised by Bethyl laboratories against a peptide pool comprising amino acids 1-20 and 176-195 of human CAS3. CAS3 is also known asFAM168B or MANI; NM_001009993). Anti-methyl CAS3R87me2a (meCAS3) was generated against the epitope CTAVYPVRme2aSAYPQQSR by Covance. Methyl MED12R1899me2a (meMED12) was custom generated against the epitope TSVYRme2aQQQP by CST. PRMT1 antibody was a gift from Stephane Richard (McGill University). H3R2me2a (Millipore, Catalog #05-808), H3R17me2a (Millipore, Catalog #07-214), H3R8me2a (Active Motif, Catalog #39651), CARM1 (Bethyl, Catalog #A300-421A) and β-actin (Sigma, Catalog #A5316) were obtained commercially.

CARM1 substrate motif antibodies were raised against an antigen mixture of six different CARM1 methylated motifs (Fig 1A), to specifically recognize endogenous proteins when asymmetrically dimethylated by CARM1. The meMED12 antibodies were raised against the peptide sequence TSVYR*QQQP of human MED12 protein (NP_005111). This work was performed in collaboration with Cell Signaling Technology (CST). mePABP1 antibody was raised against the peptide sequence CGAIR*PAAPR*PPFS of human PABP1 protein (NP_002559) (Cheng & Bedford, 2011 (link)). R* denotes asymmetrically dimethylated arginine residue. The MED4, MED30, and CDK8 antibodies were a gift from Thomas Boyer (University of Texas Health Science Center at San Antonio). The CARM1 antibody, used for ChIP-seq, was a gift from Stéphane Richard (McGill University). The following antibodies were obtained commercially: H3R17me2a (07-214; Millipore and ab8284; Abcam), MED12 (A300-774A; Bethyl), SRC-1 (2191; CST), SRC-3/AIB1 (611105; BD Transduction), CARM1 (A300-420A; Bethyl), PRMT1 (A300-722A; Bethyl), PRMT6 (A300-929A; Bethyl), CDK8 (SC-1521; Santa Cruz), and FLAG (F7425 [rabbit IgG] and F3165 [mouse IgG]; Sigma-Aldrich).