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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 2. Induction of autophagy in primary human alveolar macrophages and murine macrophages RAW264.7 cells by LPS Stimulation.

Figure 2

Cells were incubated the in the absence (control cells) or presence of LPS (100 ng/ml) or in the presence of LPS plus PMB (25 μg/ml) for 16 h. (A) Upper panel- Representative immunofluorescence images with LC3 antibody staining in RAW264.7 cells. Middle panel- Quantitation of the percentage of cells with autophagosomes. Lower panel- Western analysis using antibodies against LC3 or β-actin. (B) GFP-LC3 fluorescence images and quantitation analyses are shown in upper and middle panels, respectively. Lower panel- Western analysis with anti-iNOS antibody. Activity of iNOS was evaluated by measuring nitrite accumulation in culture media. (C) Upper panel- Representative immunofluorescence images of LC3 antibody staining in primary human alveolar macrophages. Lower panel-Quantitation of the percentage of cells with autophagosomes. (D) Ultrastructural analysis of LPS-induced autophagy by transmission electron microscopy in RAW264.7 cells. av, autophagic vacuole; n, nucleus; m, mitochondria. Graph represents quantitation of the number of autophagosomes per cross-sectioned cell. Data are mean±SEM from three (A-B) or two (C-D) independent experiments. Scale bar, 10 μm. * and ** denote p < 0.05 and p < 0.001, respectively, when compared to control condition.