Degrasyn wp1130

Degrasyn (WP1130) and PR-619 were purchased from Selleck Chemicals (Houston, TX, USA). FBS, L-glutamine, penicillin and streptomycin solution, 0.25% trypsin EDTA, trypan blue, poly-L-lysine, ribonuclease A, propidium iodide (PI), RIPA buffer, SigmaFAST Protease Inhibitor Cocktail, and TBST buffer were acquired from Sigma-Aldrich (Steinheim, Germany). Annexin V-FITC was obtained from Immunostep (Salamanca, Spain). The CellEvent^® Caspase3/7 Green Flow Cytometry Assay by ThermoFisher (Waltham, MA, USA) was used. Anti-USP5 (sc-390943), anti-USP9X/Y (sc-365353), anti-USP14 (sc-515812), anti-Bcl-2 (sc-7382), and anti-β actin (sc-47778) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), whereas the anti-Bcl-xl (#2764) antibody was purchased from Cell Signaling (Cell Signaling Technology, Danvers, MA, USA). The anti-γH2A.X (ab26350) antibody was from Abcam (Cambridge, UK), and anti-mouse/HRP and anti-rabbit/HRP antibodies were bought from Dako (Glostrup, Denmark).

Primary antibodies were obtained from the following sources: Usp9X (Cell Signaling, Danvers, MA #5751), Mcl-1 (Cell Signaling, Danvers, MA #5453), Noxa (Calbiochem, San Diego, CA #OP180), ATF4 (Cell Signaling, Danvers, MA #11815), ATF3 (Novus Biologicals, Littleton, CO #NBP1-85816), β-actin (Sigma, St. Louis, MO #A5316). Secondary antibodies were HRP-conjugated goat anti-rabbit (Thermo Scientific, Waltham, MA #31460) and goat anti-mouse (Thermo Scientific, Waltham, MA #31430). Degrasyn [WP1130] was obtained from Selleckchem (Houston, TX #S2243). A-1210477 was purchased from APExBIO (Houston, TX #B6011). Tunicamycin [TUN] and Staurosporine [STS] were purchased from Sigma (St. Louis, MO; TUN #T7765, STS #S5921).