Cellular ESCRT components are recruited to regulate the endocytic trafficking and RNA replication compartment assembly during classical swine fever virus infection
Fig 5
Interaction between major subunits of the ESCRT-0/I/II system and CSFV proteins.
(A, D and G) PK-15 cells were transfected with indicated plasmids (pFlag-Core, -E2, -NS3, -NS4B, -NS5A, -NS5B) for 48 hpt, then fixed and subjected to immunofluorescent by using rabbit anti-HRS (A); -VPS28 (D); -VPS25 (G) antibody (green) and mouse anti-Flag antibody (red). The nuclei were stained with DAPI. Bars = 10 μm. These data are representative of three independent experiments. (B, E and H) The colocalization analysis was expressed as Pearson’s correlation coefficient, respectively. Results are represented as the mean + SD of data from three independent experiments. **, P < 0.01. (C, F and I) HEK-293T cells were transfected with indicated plasmids (pFlag-NS3, -NS4B, -NS5A, -NS5B, -Core, -E2) or vector for 48 hpt, then harvested for immunoprecipitation by using mouse anti-Flag antibody or mouse anti-HRS (C); -VPS28 (F); -VPS25 (I) antibody, and whole-cell lysates were subjected to Western blotting by using rabbit anti-HRS/VPS28/VPS25 antibody or mouse anti-Flag antibody, along with β-actin as a loading control. These data are representative of three independent experiments.