Cellular ESCRT components are recruited to regulate the endocytic trafficking and RNA replication compartment assembly during classical swine fever virus infection
Fig 2
ESCRTs machinery participates in CSFV endocytosis.
(A-D) PK-15 cells were infected with CSFV (MOI = 10) at 4°C for 1 hpi, then shifted to 37°C for indicated time points, respectively. After fixed and subjected to immunofluorescent by using mouse anti-CSFV E2 monoclonal antibody (WH303) and rabbit anti-VPS25 (A); -CHMP4B (B); -CHMP7 (C); -VPS4A (D) antibody. The nuclei were stained with DAPI. The white arrow indicates the co-localized protein. Bars = 10 μm. These data are representative of three independent experiments. (E-G) PK-15 cells were infected with CSFV (MOI = 10) or not for 6 hpi, then harvested for immunoprecipitation by using mouse anti-VPS25 antibody (E) or rabbit anti-CHMP4B (F); -CHMP7 antibody (G), and the whole-cell lysates were subjected to Western blotting by using the antibodies against Clathrin, Rab5, Rab9, LAMP-1, Tsg101, VPS25, CHMP4B, CHMP7, VPS4A and ALIX. These data are representative of three independent experiments.