Fig. 3.

Mycobacterial infection further increased autophagy in TSC1 KO BMMϕ. a Tsc1 KO macrophages have mTORC1-dependent inhibition of ULK1 at S757. BMMϕ cells from either TSC1 WT or KO BMMϕ were left uninfected or infected with M. tuberculosis strain H37Rv (TB) (a, b, c) or BCG (d, e, f) at a MOI of 10 for 24 h and lysed for immunoblotting analysis with the indicated antibodies. TSC1 WT and KO BMMϕ were treated with 100 ng/ml rapamycin (b, d) or with 100 nM bafilomycin A1 (e, f) for 3 h and then lysed for Western blot analysis. c, f Densitometric quantification of LC3-II (n = 3) was normalized to total Actin in c and e. The arrow indicates LC3B-II. These experiments were repeated three times. BafA1, bafilomycin A1; BCG, Bacillus Calmette-Guérin; Rapa, rapamycin; TB, M. tuberculosis strain H37Rv, WT, Wild type; KO, knockout; UN, uninfected. *, p < 0.05; **, p <0.01