Fig. 4. Knockdown of p65 decreases Fas expression and reduces TNFα-mediated sensitization to FasL-induced apoptosis.

a RT-qPCR analysis of Fas mRNA of Hepa1-6 cells stably expressing Renilla or p65 shRNAs, untreated or treated with TNFα for 2 h. b Anti-p65 and anti-Fas western blot analysis of wild-type (Wt), shRenilla or shp65 Hepa1-6 cells, untreated or treated with TNFα for 14 h. Tubulin serves as loading control. c The same analysis as in b but in AML12 cells. FACS analysis of Fas cell surface expression on shRenilla or shp65 Hepa1-6 (d) or AML129 (e) cells, untreated or treated with TNFα for 14 h. Caspase-3/-7 activity in total cell lysates of shRenilla or shp65 Hepa1-6 cells (f) or AML12 cells (g). a.u. arbitrary units. (a, d-g) Values represent the means of at least three independent experiments ± SD