Figure 4

Parkin is required for phagolysosome formation and effective killing of bacteria. (a) Representative images show mitochondrial network in Park2^+/+ and Park2^−/− macrophages. Mito-Tracker (green), nuclei (blue). Scale bar 10 µm. (b) Images, 3-D profiles, and Mito-SOX quantitation showing Mito-SOX fluorescence in Park2^+/+ and Park2^−/− macrophages. Mito-SOX (red), nuclei (blue). Scale bar 50 µm. (c) Representative plot shows OCR indices (Mitochondrial Stress Test) in Park2^+/+ and Park2^−/− macrophages. (d) Representative Western Blot (left) of Parkin in lungs of Park2^+/+ and Park2^−/− mice. (Right) CFUs from lung homogenates of Park2^+/+ and Park2^−/− mice that were subjected to bacteria (i.t.). Mean ± s.e.m., n = 5–6 mice per group. *P < 0.05, Student’s t-test. (e) CFUs after incubation of Park2^+/+ and Park2^−/− macrophages or neutrophils with bacteria. Mean ± s.e.m., n = 4–5 individual measurements using cell populations obtained from 2 to 3 mice. *P < 0.05, Student’s t-test. (f) Western blot analysis of LC3BII/I in macrophages-treated with FCCP (0 or 500 nM) for indicated time. Mean ± s.e.m., n = 4. *P < 0.05, ANOVA. (g) Images depicted phagolysosome formation in Park2^+/+ and Park2^−/− macrophages incubated with bacteria (red). LysoTracker (green), nuclei (blue). Scale bar 10 µm.