Figure 8. Control of SIRT1 gene silencing with SIRT1-siRNA and its impact on the expression patterns of targeted epi-marks H3k4ac, H3k9ac and H4k16ac in-vitro.

(A) MCF-7, T-47D, MDA-MB 453, MDA-MB 231 and MDA-MB 468 cells were transfected with SIRT1-siRNA (siSIRT1) or negative control siRNA (Ctrl). After 48 hours of transfection, equal amounts of proteins were immunoblotted with anti-SIRT1 Ab (120 kDa), anti-H3k4ac Ab (17 kDa), anti-H3k9ac Ab (23 kDa) and anti-H4k16ac Ab (27 kDa). β-actin (42 kDa) served as an internal loading control. (B) Relative expression levels were evaluated using Quantity One software and normalized against the internal control β-actin. Each bar represents the percentage contribution of each of the 4 proteins compared to the total set as (100%). All experiments were performed in triplicate fashion.