Inhibition of autophagy by pharmacological inhibitors reduced LPS-induced bactericidal activity. HMrSV5 cells were treated for 12 hours in the absence (control) or presence of LPS (1.0 μg/ml), DMSO, 3-MA (10 mM), Wm (50 nM), LPS + 3-MA or LPS + Wm. (A) The panel shows a western blot probed with antibodies against Beclin-1, LC3-II or β-actin. (B) Densitometric analysis of Beclin-1 or LC3-II in Figure 5A; β-actin was used as a loading control. (C) Autophagic vacuoles were labeled with MDC (blue) in the left panel. Scale bars: 20 μm. The graphs on the right panel represent quantitation of the number of MDC-labeled autophagosomes per cell. *p < 0.05 in Figure 5B (vs. control); ** p < 0.01 in Figure 5C (vs. control); # p <0.05 in Figure 5B and 5C (vs. LPS) (D) Graphs represent percentage of remaining E.coli in each group as described above. Data represent mean values ± SD (n ≥ 3). * and ** denote p < 0.05 and p < 0.01 respectively (LPS vs. control); # and ## denote p < 0.05 and p < 0.01 respectively (LPS + 3MA or LPS + Wm vs. LPS).