SNX8 is essential for VPS34-mediated trafficking of MITA.

(A) HEK293 cells (2x106) were transfected with HA-SNX8 and the indicated plasmids (5μg each) for 24 h before co-immunoprecipitation and immunoblot analysis were performed. (B) HEK293 cells (2x106) were transfected with Flag-VPS34 (5 μg) and the indicated plasmids (5 μg each) for 24 h before co-immunoprecipitation and immunoblot analysis were performed. (C) HeLa cells (1x105) were transfected with HA-VPS34 (0.2 μg) and GFP-tagged SNX8 (0.1 μg) and MITA (0.1 μg) for 20 h before immunostaining was performed with anti-HA. (D) MLFs (3x107) were left un-infected or infected with HSV-1 (MOI = 1) for the indicated times before endogenous co-immunoprecipitation and immunoblot analysis were performed. (E) HEK293 cells (2x106) were transfected with the indicated plasmids (5 μg each) for 24h before co-immunoprecipitation and immunoblot analysis were performed. (F) SNX8+/+ and SNX8-/- HeLa cells (1x105) were transfected with HA-VPS34 (0.2 μg) and GFP-tagged MITA (0.1 μg) for 20 h before immunostaining was performed with anti-HA. (G) MEFs cells (2x105) were transfected with VPS34 siRNA (2 μg) for 36 h and then infected with HSV-1 (MOI = 1) for 3 h. Immunostaining were performed with the indicated antibodies. (H) VPS34-RNAi stable-knockdown THP1 cells (4x105) were infected with HSV-1 (MOI = 1) for 6 h before qPCR analysis. (*p<0.05, **p<0.01).