Figure 5

AMPK activation restored autophagy and bacterial killing in Parkin-deficient macrophages. (a) Western blot analysis of phospho AMPK and ACC in AICAR (0 or 500 µM) treated Park2^−/− macrophages, while (b) shows Parkin levels after sequential exposure to AICAR (0 or 500 µM) for 2.5 h and LPS (0 or 300 ng/ml) for 24 h. Mean ± s.e.m., n = 3–4. *P < 0.05, Student’s t-test; ^#P < 0.05, ANOVA; ns, not significant. (c) LC3BII/I ratios in Park2^−/− macrophages-treated with AICAR (500 µM) for indicated time. Mean ± s.e.m., n = 3. *P < 0.05, ANOVA. (d) Representative Western Blots and optical bend densitometry of pS93-Beclin-1 in Park2^−/− macrophages-treated with or without AICAR (500 μM) for 4 h. Mean ± s.e.m., n = 4. *P < 0.05, Student’s t-test. (e) The amounts of CFUs after incubation of Park2^+/+ and Park2^−/− macrophages with bacteria. Park2^−/− macrophages were also pre-treated with AICAR (0 or 500 µM). Mean ± s.e.m., n = 5. ^#P < 0.05, ANOVA. (f) The effects of direct activators of AMPK on bacterial killing by peritoneal macrophages ex vivo. PAK CFUs are determined from Park2+/+ and Park2−/− macrophages that were treated with or without MK8722 (0.6 µM) or A769662 (10 µM) for 60 min prior to exposure to bacteria. Data are presented as Box Plot, n = 8–10. *P < 0.05, ANOVA.