FIGURE 3.

NOD1 and NOD2 are required for autophagy activation upon L. monocytogenes infection. A, Western blot analyses of LC3-II in WT, Nod1^−/−, Nod2^−/−, or Rip2^−/− macrophages that were uninfected (C), infected with L. monocytogenes (Lm) for different periods (in the presence or absence of CQ), or incubated with rapamycin (Rap) (50 μg/ml) for 2 h. Actin was used as a loading control. B, densitometry scanning of the blot showing the ratio of LC3-II to actin in wild-type, Nod1^−/−, Nod2^−/−, or Rip2^−/− macrophages at 4 h. The graph is a representation of three different blots. C, confocal microscope images of WT, NOD1-, NOD2-, or RIP2-deficient macrophages transfected with the GFP-LC3 plasmid. Cells were uninfected, infected with L. monocytogenes for 4 h, or treated with rapamycin (50 μg/ml) for 2 h. Scale bars = 10 μm. D and E, quantification of the number of GFP-LC3 puncta per cell at 4 h post-infection in Nod2^−/− macrophages or Rip2^−/− macrophages (E) when infected as described in C. Results show mean ± S.E. (*, p ≤ 0.05) of experiments done at least three times.