Anti p19arf

Protein lysates were prepared in RIPA buffer and 20–50 μg of protein were fractionated in a 10 % SDS-PAGE gel and transferred to a PVDF membrane (Immobilon-P, Millipore). The membrane was blocked with 5 % milk (Bio-Rad Laboratories), probed with antibodies, followed by washing and incubation with HRP-conjugated secondary antibody (Pierce) and developed using the WesternBright ECL^TM reagent (Advansta, USA). Antibodies used were anti-V5 (Life Technologies, USA), anti-p53 (Becton Dickinson, USA), anti-Myc, anti-p53-Ser15, anti-p21 (Santa Cruz Biotechnology, USA), anti-Caspase3, anti-PARP, anti-Bmi1, anti-Bcl2, anti-Bcl-xL, anti-Snail, anti-Slug (Cell Signaling, USA), anti-Bcl-W (GeneTex, USA), anti-p19ARF, anti-JMJD6 (Abcam, USA), anti-vimentin, and anti-β-actin (Millipore, USA).

As described previously [10 (link)] mucosal stripping were obtained from the TgM9 and WT mice (n = 20 per group) with and without CAC after the sacrifice for WB analysis. The antibodies used were anti-MMP9 (Abcam, Cambridge, MA), anti-NICD (Abcam), anti-caspase-3 (Cell Signaling, Beverly, MA) anti-p53 (Cell Signaling), anti-p21^WAF1/Cip1 (BD Bioscience, San Jose, CA), anti-cyclin D1 (Santa Cruz, Dallas, TX), anti-Cyclin A (Cell Signaling), anti-Cyclin E1 (Santa Cruz), anti-γH2AX (Abcam), anti-p19ARF (Abcam), and anti-p14ARF (Abcam). Goat anti-mouse secondary antibody (1:2000; Bio-Rad, Hercules, CA) or goat anti-rabbit secondary antibody (1:2000, Bio-Rad) were used. Densitometry graphs were generated by using image acquisition and analysis software by VisionWorksLS Analysis Software (UVP, Upland, CA)