Fig. 6

Downregulation of autophagy in TP53-rescued and SAHA-treated ESS-1 cells demonstrated by MDC/PI staining. a MDC (green) and PI staining (red) was performed of TP53-WT, TP53-637C>T, or non-transfected ESS-1 cells treated with 3 µM SAHA to quantify autophagosome formation and cytotoxicity, respectively. Untreated, SAHA, SAHA and TRAIL-treated, as well as TP53-WT-transfected cells were included as controls. Rapamycin-supplied cells were included as a reference for induction of autophagy. TP53-WT-rescued cells pretreated with 10 µM caspase family inhibitor (CFi) served as a control to monitor the influence of caspase-dependent apoptosis. b Representative confocal images used for quantitative autophagic and cytotoxicity measurements in a. At least three different images resulting from three independent experiments were used for analysing MDC (upper panel; green fluorescence) and PI (lower panel; red fluorescence) staining. The scale bar indicates 50 µM (Magnification: 40-fold)