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. Author manuscript; available in PMC: 2009 May 12.
Published in final edited form as: Immunity. 2007 Jul 19;27(1):135–144. doi: 10.1016/j.immuni.2007.05.022

Figure 5. Inhibition of PI3-kinase class III activity blocks LPS-induced autophagy.

Figure 5

(A) RAW264.7 cells were incubated for 16 h in the absence (control), or presence of LPS, LPS plus 3MA (5 mM) or LPS plus Wortmannin (Wm, 100 nM). (B) RAW264.7 cells stably expressing GFP-LC3 were subjected to the same conditions as in (A). Cells were fixed, stained with DAPI to visualize the nuclei (blue). (C) RAW264.7 cells stably expressing a dominant negative (DN) mutant of VPS34 were incubated with LPS for 16 h in parallel with RAW264.7 cells incubated with or without LPS. Autophagic vacuoles were stained in fixed cells using MDC (blue). Deconvolution microscopy images are shown. Scale bar, 10 μm. Graphs represent quantitation analysis of the number of MDC-positive autophagosomes per cell (A and C) or the percentage of cells with GFP-LC3-positive autophagosomes (B). Data represent mean ±SEM of three independent experiments. * and ** denote p< 0.05 and p< 0.001, respectively, when compared to control condition.