Image studio lite western blot analysis software

The proteins were extracted from the cells or colonic biopsies by RIPA buffer (Sigma, St. Louis, MO, US). Extract proteins (50 μg) were separated by 12% SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane, probed with 1:1000 dilution of primary antibodies (below), and incubated with FITC-conjugated anti-mouse or anti-rabbit immunoglobulin (1:15000 dilution; Millipore, MA, USA). Primary antibodies used for Western blot analyses include: Anti-hMLH1, -hMSH2, and -ERβ, polyclonal antibodies were purchased from Epitomics (Burlingame, CA, USA); anti-caspase 3 and -caspase 9 were purchased from Cell Signaling Technology (Beverly, MA, USA); anti-Bax, -P53 and -GAPDH monoclonal antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Expression of proteins of interests was imaged using an ODYSSEY quantitative fluorescent imaging system (LI-COR, Lincoln, NE, USA), and quantified by Image StudioTM Lite Western Blot Analysis software (LI-COR, Lincoln, NE, USA).

Levels of proteins of interest evaluated by Western blot were quantified using Image StudioTM Lite Western Blot Analysis software (LI-COR, Lincoln, NE, USA) and normalized to GAPDH; and statistical analysis between groups was performed using ANOVA. The SPSS 13.0 software was used for statistical analyses of FACS data. P value <0.05 was considered statistically significant. At least 3 independent experiments were performed in each set of data presented.