FIG 5.

The autophagy-related proteins LC3 and p62 do not localize to PVs in alveolar epithelial cells. (A) THP-1 and A549 cells were infected with C. burnetii for 72 h. Cells were processed for fluorescence microscopy using antibodies against LC3 (green), p62 (green), CD63 (red), and C. burnetii (violet), with DAPI to stain nuclei (blue). Images are representative of three separate biological experiments. Bars, 20 μm. THP-1 and H292 cells and HLFs contain PVs that are decorated with LC3 and p62, while PVs in A549 cells are devoid of both proteins. (B) The colocalization of LC3 or p62 with PVs in each cell type at 72 hpi was quantified and averaged. ***, P < 0.001; ****, P < 0.0001; n.s., not significant. A549, H292, and HLF data were compared to THP-1 data for statistical analysis. PVs in A549 cells are devoid of LC3 and p62, whereas PVs in HLFs are not significantly different from those in THP-1 cells.