Time- and concentration-dependent stimulation of autophagy by rapamycin.

<p>A–B) Western-blot analysis for GAPDH and LC3 of protein lysates obtained from HeLa cells treated with DMSO or 1 µM rapamycin (Rapa) for the indicated time periods (A) (<i>n</i> = 7) or for 5 h with the indicated concentrations (B) (<i>n</i> = 6). One hour before harvesting, 100 nM bafilomycin A1 was added. Upper panels: representative Western blots; lower panels: quantification of the LC3-II/GAPDH ratio. C) GFP-LC3-punctae quantification in HeLa cells treated for 5 h with different concentrations of rapamycin. Left: representative pictures. The scale bar represents 10 µm. Concentrations are mentioned in the right lower corner. Right: Quantification of the number of punctae per cell (<i>n</i> = 3). * <i>p</i><0.05, repeated measurements ANOVA.