Fig. 3: Autophagy stimulation reduces apoptosis in starved CSCs.

A Experimental timeline and design for autophagy modulation and cell viability assessment. B Representative contour plot of the positive control of apoptotic induction. Representative flow cytometry density plots (C) and relative quantification (D) of early (Annexin-V+/7AAD-) and late (Annexin-V+/7AAD+) apoptotic CSCs after 32 hours of nutrient deprivation (0.1% FBS, 0 mM glucose), with or without pretreatment with 50 mM trehalose (TRE) for 24 hours (n=8); corresponding WB panel for cleaved caspase 3 is shown in paneI (E). Representative flow cytometry density plots (F) and relative quantification (G) of early and late apoptotic cells after 32 hours of nutrient deprivation in CSCs that were previously transduced with ad-ATG7 or ad-LacZ 48 hours before stress (n=7); corresponding WB panel for cleaved caspase 3 is shown in panel (H). Representative flow cytometry density plots (I) and relative quantification (J) of early and late apoptosis after 32 hours of nutrient deprivation in CSCs that were pre-treated with si-ATG7 or scramble control (Scr) transfection 48 hours before stress (n=7); corresponding WB panel for cleaved caspase 3 is shown in panel (K). *P<0.05, **P<0.01, ***P<0.001.