Figure 7. Knockdown of WIPI1, WIPI2, ATG5 and ATG7 but not BECN1 attenuated rVP1-mediated MAPK1/3 phosphorylation and MMP9 activity. (A) RAW264.7 cells transfected with scrambled or target siRNA as indicated were incubated with or without 4 μM rVP1 for 4 h. Cell lysates were collected and subjected to immunoblot analysis using antibodies against LC3, phosphorylated AKT1 at Ser473, phosphorylated RAF1 at Ser338, phosphorylated MAPK1 (Thr185/Tyr187), phosphorylated MAPK3 (Thr202/Tyr204) and their nonphosphorylation controls. ACTB was used as a loading control. Blots are representative of three independent experiments. (B) Knockdown of WIPI1, WIPI2, ATG5 and ATG7 but not BECN1 attenuated rVP1-induced MMP9 activity. Cells transfected with scrambled or target siRNA as indicated were incubated with or without 4 μM rVP1 for 24 h. Supernatants of cells were collected for examine MMP activity by gelatin zymographic analysis. Data represent means ± SEM of three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001, N.S., not significant.