Figure 5

IRF-1 inhibits autophagy through iNOS/mTOR/P70S6 in vitro. (A) AdIRF-1 infection induced IRF-1 overproduction. AdLacZ and AdIRF-1 infected RAW 264.7 cells for 24 h at either an MOI of 100 or 200 and Western blot analysis for IRF-1 expression was performed. (B) RAW cells were infected with AdIRF-1 at an MOI of 200 for 24 h; cells were then harvested for iNOS expression. Griess assay confirmed NO production following AdIRF-1 infection. (C) Peritoneal macrophages were isolated from WT and IRF-1 KO mice and stimulated with LPS (1 µg/mL) for 16 h and analyzed by Western blot for iNOS, p-mTOR and p-P70S6. (D) RAW cells were infected with AdIRF-1 at an MOI of 200 for 24 h and were treated with L-NIL (200 µmol/L) for 16 h. Cells were then harvested for Western blot of p-mTOR, p-P70S6, LC3B I/II and P62. *^,#P < 0.05. Results representative of three separate independent experiments.