Figure 3

From: iASPP facilitates tumor growth by promoting mTOR-dependent autophagy in human non-small-cell lung cancer

Figure 3

shiASPP-induced autophagy inhibition is dependent on mTOR activation in A549 cells. (a) Western blot analysis of mTOR pathway proteins in scramble or shiASPP cells, either with iASPP rescue or with rapamycin treatment. (b) Endogenous levels for LC3 (red fluorescence) were visualized on confocal microscope by immunofluorescenc staining. (c) LC3 dots in scramble control or shiASPP cells treated with mTOR inhibitor as in (b) were counted. Data in (c) represent mean±S.E.M. of at least 100 cells scored (**P<0.01). (d) Conversions of LC3-I to LC3-II in cells treated with rapamycin were determined by western blotting. GAPDH was a loading control. (e) The ratio of endogenous LC3-II to LC3-I protein level treated as in (d) was counted. The value in the scramble control cells with rapamycin treatment was normalized as 1. (f) Cells were treated with rapamycin at different doses, and their viability was determined by the CCK8 assay (*P<0.05; ***P<0.001; NS, no significance)