Figure 7

Restoring NF-κB activity attenuates the suppressive effect of Hsp27 knockdown in ALDH+ breast cancer cells. AS-B145 or AS-B244 cells were transfected with negative control siRNA (Ctrl-siRNA, 100 nM), ctrl-siRNA (50 nM)+si-Hsp27 (50 nM) or si-Hsp27 (50 nM) +si-IκBα (50 nM) for 48 h. (A) The expression of Hsp27 and IκBα was determined by Western blot. Inserted values indicated relative expression in comparison with ctrl-siRNA. (B) The NF-κB activity was determined by co-transfected with NF-κB-Luc reporter vector (0.5 μg) and RLuc (0.05 μg) vector. The results were presented as relative luciferase activity to ctrl-siRNA. (C, D) The percentage of ALDH+ population among groups was determined by ALDEFLUOR assay. DEAB was used to set the cutoff line for ALDH+ cells. The quantitative results were presented as relative percentage of negative control siRNA (ctrl-siRNA) group and were shown in bar graph (D). *, P < 0.05; #, P < 0.01. ALDH, aldehyde dehydrogenase; NF-κB, nuclear factor kappa B.