Anti histone h2b

For western blots, protein extracts were prepared by washing and detaching ESCs as previously mentioned. Protein was extracted using RIPA buffer (50 mM Tris-HCl pH:8.8, 150 mM NaCl, 0.1% sodium dodecyl sulphate (SDS), 0.5% deoxycholate and 0.5% NP-40) and the concentrations quantified using the BCA Protein Assay kit (Thermo). Protein were resolved on 10% SDS-polyacrylamide gels and transferred to polyvinylidene difluoride (PVDF) (Millipore) for immunoblot detection. Primary antibodies for all the polyadenylation proteins were purchased from Bethyl Laboratories (Montgomery, TX) with the exception of CstF-64 (3A7) and τCstF-64 (6A9), which were used as described (21 (link)). Other antibodies used were rabbit polyclonal anti-FLASH (Millipore), anti-SLBP (Cell Signaling), anti-Histone H2B (Cell Signaling), anti-Cyclin A (Santa Cruz), anti-CDK2 (Cell Signaling) and anti-Histone H3 (Cell Signaling); and mouse monoclonal anti-Cyclin B1 (Millipore), anti-Histone H4 (Cell Signaling), anti-CDK4 (Cell Signaling) and anti-FLAG (Sigma).

Anti-Mov10 antibodies (ab80613) were from Abcam (Cambridge, MA, USA). Anti-PORCN antibodies (HPA049215) were from Sigma (St Louis, MO, USA). Anti-FLAG M2 antibodies (F3165) were from Sigma. Anti-HA antibody (MMS-101R) was from Covance (Conshohocken, PA, USA). Anti-Wnt5a (AF645) and anti-Ror2 (AF2064) were from R&D Systems (Minneapolis, MN, USA). Anti-histone H2B, anti-Histone H3, anti-β-actin, anti-β-tubulin and anti-GAPDH antibodies were from Cell Signaling Technologies (Danvers, MA, USA). Anti-FASN (610962) antibodies were from BD Transduction Laboratories (Franklin Lakes, NJ, USA). FASN inhibitor, Cerulenin (C2389), was from Sigma. SCD inhibitor, CAY10566 (10012562), was from Cayman Chemical (Ann Arbor, MI, USA).