Figure 10.

Inhibition of ATG13-dependent autophagy suppresses Staphylococcus-induced cell death. A, ATG13 was targeted in HEK293A cells using the CRISPR–Cas9 system (CR–ATG13). WT HEK293A or CR–ATG13 cells were treated to bafilomycin A1 (Baf) (10 nm) ± starvation (St) in EBSS for 2 h. Cell lysates were analyzed for LC3B lipidation as in Fig. 1. n = 2 ± range. B and C, WT HEK293A or CR–ATG13 cells were starved in EBSS for 2 h, fixed, and stained to detect LC3B puncta. Average puncta/cell were from n = 36 cells ± S.D. Scale bar, 10 μm. D, WT HEK293A or CR–ATG13 cells were infected with varying multiplicities of infection of S. aureus NCTC8325 and assayed for cell viability as in Fig. 6. **, p < 0.001; *, p < 0.01 by unpaired t test: CR–ATG13 versus 293A (comparing equivalent multiplicities of infection). E, WT HEK293A or CR–ATG13 cells were infected with 1:100 diluted S. enterica sv. typhimurium and incubated for 24 or 48 h before staining for cell viability. Averages are from n = 3 ± S.D. a, p = 0.054 by t test: infected CR–ATG13 versus 293A at 24 h.