Fig. 7

NS38 manipulates p62 to degrade N protein and affects biological function of N protein. A, B NS38 associates with autophagy receptor p62. EPC cells seeded in 10-cm² dishes overnight were transfected with the indicated plasmids (5 μg each). After 24 h, cell lysates were immunoprecipitated (IP) with anti-Flag affinity gels or anti-HA affinity gels. Then, the immunoprecipitates and cell lysates were analyzed by immunoblotting (IB) with anti-Flag, anti-Myc and anti-HA antibodies (Abs), respectively. C, D The p62 is essential for NS38 degradation of N protein. Overexpression of p62 promotes the degradation of N protein by NS38 (C). EPC cells were seeded in 6-well plates overnight and co-transfected the indicated plasmids. After 24 h, the cell lysates were subjected to IB with indicated Abs. Knockdown of p62 eliminates the degradation of N protein by NS38 (D). EPC cells were seeded in 6-well plates overnight and transfected with 2 μg shp62#1-pLKO.1, shp62#2-pLKO.1 or the negative control (sh-NC) with other indicated plasmids for 24 h. The cell lysates were then subjected to IB with indicated Abs.