Anti poly adp ribose polymerase 1 parp 1

Anti-vimentin, anti-Snail, and anti-Slug antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-NUCB2, anti-Twist1, and anti-Ki-67 antibodies were obtained from Abcam (Cambridge, MA, USA). Anti-E-cadherin, anti-p21^waf1, anti-BCL2, and anti-cyclin D1 antibodies were from Dako (Copenhagen, Denmark). Anti-p27^kip1, anti-Rb, anti-X-linked inhibitor of apoptosis (XIAP), anti-BAX, and anti-N-cadherin antibodies were from BD Biosciences (San Jose, CA, USA). Anti-ZEB1 and anti-β-actin antibodies and adriamycin (ADR: D1515) were from Sigma-Aldrich Chemicals (St. Louis, MO, USA). Anti-phospho (p) Rb at Ser807/811, anti-cleaved caspase-3, and anti-poly (ADP-ribose) polymerase 1 (PARP1) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-cyclin A2 and anti-cyclin B1 antibodies were from Novocastra (Newcastle, UK) and Santa Cruz Biotech (Santa Cruz, CA, USA), respectively.

Kidney tissues were prepared using a lysis buffer (20 mM Tris-HCl (pH 7.4), 1% Nonidet P-40 (NP-40), 5 mM ethylenediaminetetraacetic acid (EDTA), 2 mM Na₃VO₄, 100 mM NaF, 10 mM Na₄P₂O₇, 100 μM phenylmethylsulfonyl fluoride (PMSF), 7 μg/mL aprotinin, and 7 μg/mL leupeptin). Proteins were resolved by sodium dodecyl sulfate- (SDS-) polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride (PVDF) membrane. After blocking, the membrane was incubated with the following primary antibodies: anti-Bax (Cell Signaling Technology, Danvers, MA, USA), anti-caspase-3 (Cell Signaling Technology), anti-poly(ADP-ribose) polymerase-1 (PARP-1; Cell Signaling Technology), anti-heme oxygenase-1 (HO-1; Santa Cruz Biotechnology), anti-NAD(P)H:quinone oxidoreductase 1 (NQO1; Santa Cruz Biotechnology), and anti-β-actin (Sigma-Aldrich). The membrane was washed and incubated with a horseradish peroxidase-conjugated secondary antibody, and signals were detected using an enhanced chemiluminescence (ECL) Western blotting detection system (ImageQuant LAS4000; GE Healthcare Life Sciences, Pittsburgh, PA, USA). Relative protein expression was quantified using NIH ImageJ software.

Anti‐Ezrin/radixin/moesin (ERM), anti‐phospho (p) Ezrin (at Thr567)/radixin (at Thr564)/moesin at Thr558) (pERM), anti‐poly (ADP‐ribose) polymerase 1 (PARP1), and anti‐vimentin antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti‐EBP50, anti‐β‐actin, anti‐MYH9 (mouse), anti‐vinculin, and anti‐Sox2 antibodies were obtained from Abcam (Cambridge, MA, USA). Anti‐E‐cadherin, anti‐p21^waf1, and anti‐cyclin D1 antibodies were from Dako (Copenhagen, Denmark). Anti‐cyclin A2 and anti‐cyclin E antibodies were from Novocastra (Newcastle, UK). Anti‐p27^kip1, anti‐aldehyde dehydrogenase 1 (ALDH1), and anti‐N‐cadherin antibodies were from BD Biosciences (San Jose, CA, USA). Anti‐MYH9 (Rabbit) and anti‐cyclin B1 antibodies were purchased from Proteintech (Rosemont, IL, USA) and Santa Cruz Biotech (Santa Cruz, CA, USA), respectively. Blebbistatin and MG132 were obtained from Toronto Research Chemicals (North York, ON, Canada) and Sigma‐Aldrich Chemicals (St. Louis, MO, USA), respectively.