Rabbit igg isotype control 31 235

The antibodies used for co-immunoprecipitation and/or western blot analyses were the following: anti-ERα C-terminal (F-10 sc-8002, Santa Cruz Biotechnology, Dallas, Texas), anti-ERα (ab3575) and anti-KMT4/Dot1L (ab72454) from Abcam (Cambridge, UK), anti-Dot1L (A300-953A), antimenin (A300–105 A) and anti-WSTF (BAZ1B; A300-446A) from Bethyl Laboratories (Montgomery, Alabama), anti-β-actin (A1978) from Sigma-Aldrich (Milan, Italy), Rabbit IgG Isotype Control (31,235) from Thermo Fisher (Milan, Italy). Compounds used for cell treatments were 4-hydroxytamoxifen (TAM; H7904, Sigma-Aldrich), fulvestrant (ICI; I4409, Sigma-Aldrich), 17β-estradiol (E887-5G, Sigma-Aldrich). Pharmacological inhibition was performed with EPZ004777 (S7353; only for RNA-seq experiments) and Pinometostat EPZ5676 (S7062; for all the other experiments included in the manuscript) for Dot1L and with MI-136 (S7815) and MI-503 (S7817) for menin, all purchased from Selleckchem (Houston, TX, USA). All inhibitors were dissolved in DMSO (Sigma-Aldrich), which was therefore used as a vehicle.

Rabbit anti-phospho-TAZ antibody was previously described (34 (link)). Other antibodies and the reagents used in this study are as follows: mouse monoclonal anti-GFP (sc-9996), mouse monoclonal anti-poly (adp-ribose) polymerase (sc-8007), mouse monoclonal anti-CAS (H-2) (sc-271537), mouse monoclonal karyopherin α1 (187.1) (sc-101292) (Santa Cruz Biotechnology); mouse monoclonal anti-TAZ (560235) (BD Biosciences); rabbit polyclonal anti-α-tubulin (PM054) and rabbit polyclonal anti-β-actin (PM053) (Medical and Biological Laboratories Co Ltd); anti-DYKDDDDK-tag (014-22383) and anti-DYKDDDDK-beads (016-22784) (Wako Pure Chemical Industries); rabbit polyclonal anti-mCherry (GeneTex); rabbit monoclonal FoxO3a (75D8)(2497) and rabbit monoclonal YAP1(D8H1X)XP (14074) (Cell Signaling Technology); rabbit polyclonal importin α5 (18137-1-AP) and rabbit polyclonal TAZ (23306-1-AP) (Proteintech Group, Inc); mouse monoclonal β-catenin (13-8400) (Zymed Laboratory Inc); Protein G Sepharose 4 Fast Flow (17-0618-01) (GE Healthcare BioSciences); Rabbit IgG Isotype Control (31235) (Thermo Fisher Scientific); and TI-4 (2,2-dichloro-N-(4-nitrophenyl)-3-phenylcyclopropanecarboxsmide) (STK825627; Vitas-M Chemical Limited).