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. 2011 Apr 14;2(4):e144. doi: 10.1038/cddis.2011.29

Figure 6.

Figure 6

SD induces autophagy in RGC-5 cells. (a) Effect of SD on Beclin-1 and LC3 expression. Representative western blot showing the expression of Beclin-1 and LC3I and II in RGC-5 cell control (10% FBS; CTR) and starved (0% FBS; SD) for 24 h. Histograms show densitometric analysis. Data were normalized for GAPDH and expressed as percentage of control values, n=5, *P<0.05. (b) Immunocytochemistry for LC3 in RGC-5 cells in presence (control) or absence of serum (serum deprived). After 24 h of SD, LC3 immunoreactivity shows a punctuate pattern due to the localization of LC3II in the autophagosome membrane. (c) SD reduces RGC-5 cell survival. Cell viability was measured by MTT assay (CTR=100±1.5; S.D.=54±1.5; n=5). (d) Effect of BafA1 treatment on LC3 expression. Western blot, representative of three independent experiments, shows the accumulation of LC3I and II in RGC-5 treated with BafA1 in presence or absence of serum for 24 h. (e) Effect of treatment with autophagy inhibitors on cell survival of starved RGC-5. RGC-5 cells were treated with BafA1 (100 nM) and 3-MA (10 mM) for 24 h under SD and viability evaluated by MTT assay (n=3). Each bar represents the mean±S.E.M. *P<0.05