Western Blot Analysis of Cancer Stemness

OVCAR3 and A2780 cell lines were processed for protein isolation and Western blotting using standard procedures, as described previously [19 (link)]. The following primary antibodies were used: anti-hPaf1/PD2, anti-Leo1, anti-Parafibromin, anti-Ctr9 (Bethyl Laboratories, Montgomery, TX, USA); anti-OCT3/4, anti-SOX-2, anti-CD24, anti-ESA, anti-SHH, anti-HER2 (Santa Cruz Biotechnology, Dallas, TX, USA); anti-CD44 (Cell signalling Technology, Danvers, MA, USA); anti-CD133 (Abnova, Walnut, CA, USA); anti–β-Catenin (Sigma); and anti-Lgr5, anti-SOX-9 (Abcam, Cambridge, MA, USA) overnight at 4°C. β-actin was used as a loading control. The band intensity was quantified using ImageJ and the normalization was performed as described previously [18 (link)].