Anti cleaved parp 9541

Antibodies against AR (sc-816), prostate-specific antigen (PSA; sc-7638) and FKBP5 (sc-11514) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). anti-HER2 (#2165), anti-phosphorylated Y-box binding protein-1^Ser102 (p-YB-1; #2900), anti-α-tubulin (#2125), anti-Akt (#9272), anti-phosphorylated Akt^Ser473 (p-Akt; #4060), anti-PARP (#9542), and anti-cleaved PARP (#9541) antibodies were obtained from Cell Signaling (Danvers, MA). Antibodies against Lamin B1 (ab16048) were purchased from Abcam (Cambridge, MA). Anti-YB-1 (2397-1) antibodies were obtained from Epitomics (Burlingame, CA). Anti-β-actin (A3854) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ) and Sigma (St Louis, MO).

To study the induction of apoptosis, cells were treated with camptothecin (Wako, Osaka, Japan), doxorubicin (LC Laboratories, Woburn, MA), MG132 (Merck Millipore, Darmstadt, Germany), or exposed to γ-irradiation (Gammacell 40, Atomic Energy of Canada, Ontario, Canada). Expression of cleaved poly (ADP-ribose) polymerase (PARP) and cleaved caspase-3 was detected by western blot analysis using anti-cleaved PARP (9541) and anti-caspase-3 antibodies (9662), respectively (Cell Signaling Technology, Danvers, MA). Assessment of apoptosis was also performed by annexin V and PI double-staining using Alexa Fluor 488 Annexin V/Dead Cell Apoptosis Kit (Life Technologies). Briefly, cultured cells were treated with vehicle or camptothecin for 24 h. The cells were stained with Annexin V and PI, and subsequently analyzed on a FACSCalibur (Becton Dickinson, Franklin Lakes, NJ) using FlowJo software (Tree Star, Ashland, OR).

Rabbit polyclonal anti-Poly ADP-ribose polymerase (PARP) (#9542), anti-cleaved PARP (#9541), and anti-cyclin B1 (#4138P) antibodies and mouse monoclonal anti-cyclin A2 (#4656P; clone BF683) and anti-p21 (#2946P; clone DCS60) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Rabbit polyclonal anti-cyclin D1 (RM9104-S1) antibodies was obtained from Thermo Fisher Scientific (Waltham, MA, USA). A rabbit polyclonal anti-zinc finger E-box binding homeobox 1 (ZEB1) (HPA028524) antibody and mouse polyclonal anti-beta actin (A5441) antibody were obtained from Sigma-Aldrich. A rabbit monoclonal anti-p27 antibody (ab32034, clone Y236) was obtained from Abcam (Cambridge, MA, USA). Mouse monoclonal anti-cyclin E1 (SC-247, clone HE12), anti-p16 (SC-1661; clone F-12), and anti-c-Myc (SC-40; clone 9E10) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). A mouse monoclonal anti-E-cadherin (610181; clone 36/E-Cadherin) antibody was obtained from BD Bioscience (Franklin Lakes, NJ, USA). A mouse monoclonal anti-vimentin (M0725, clone V9) antibody was obtained from DAKO (Glostrup, Denmark). JQ1 (1268524-70-4) was obtained from Cayman Chemical (Ann Abor, MI, USA). Lentiviral sh-RNA vectors were from Dharmacon (Lafayette, CO, USA).