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. 2015 Jan 28;10(12):2193–2207. doi: 10.4161/15548627.2014.981786

Figure 7.

Figure 7.

Autophagy is essential for MTORC2 activity induced by serum starvation. (A) Upper panel: Evaluation of AKT Ser473 phosphorylation (AKT p) and total AKT (AKT t) by WB in WI-38 fibroblasts at baseline or exposed to SS plus vehicle for 2 d or maintained in SS with the autophagy inhibitors 3-methyladenine (1 mM; 3-MA), wortmannin (100 nM; W), or LY294002 (5 μM; LY). Representative of 4 independent experiments. Lower left panel: Densitometric analysis of AKT p relative to AKT t in cells exposed to DMSO or 3-MA (representative of 4 independent experiments, *p = 0.0286). Lower right panel: Densitometric analysis of AKT p relative to AKT t in cells exposed to DMSO or LY (representative of 4 independent experiments, *p = 0.0114 V vs LY at d 2). (B) Upper panel: Evaluation of AKT Ser473 phosphorylation (AKT p) and total AKT (AKT t) by WB in WI-38 fibroblasts exposed to SS for 2 d post-transfection with control siRNA (siCTL) or siRNA specific to ATG7 (siATG7). Representative of 3 independent experiments. Lower panel: Densitometric analysis of AKT p protein level relative to AKT t (representative of 3 independent experiments, ***P < 0.0001) in WI-38 fibroblasts silenced for ATG7 expression (ATG7 silencing is effective at 80.6% ± 6.0%, representative of 3 independent experiments, ***P < 0.0001).