Phospho-Vimentin (Ser39) Antibody - #AF8233

FIG 3 VIM rearrangement is regulated by ROCK during CSFV infection. (A) CSFV replication promoted ROCK expression that induced phosphorylation of VIM at Ser72. Cells were infected with CSFV (MOI = 1) at 37°C. At indicated time points of the infection, levels of protein expression of p-(ser72)VIM, p-(ser39)VIM, VIM, and ROCK were measured by Western blotting. (B) Y27632 inhibited phosphorylation of VIM at Ser72. Cells treated with different concentrations of Y27632 were infected with CSFV (MOI = 1). At 24 hpi, cells were harvested and subjected to Western blotting. (C) The treated cells above were fixed and stained with mouse anti-E2 antibody (green) and observed by confocal microscopy. Bars = 50 μm. (E) The cytotoxic effect of thiazovivin on cells was evaluated by CCK8 assay. (F) Cells treated with different concentrations of thiazovivin were inoculated with CSFV (MOI = 1) and harvested for RT-qPCR and virus titration at 24 hpi. The bars and line symbols indicate viral mRNA copy numbers and virus titers in the panel, respectively. (G and H) Cells treated with different concentrations of thiazovivin were infected with CSFV (MOI = 1) at 37°C. At 24 hpi, cells were harvested and subjected to Western blotting and confocal microscopy. Bars = 50 μm. (D and I) The fluorescence intensity of CSFV E2 is expressed as relative fluorescence intensity. (J) Y27632 and thiazovivin inhibited VIM rearrangement. Cells treated with Y27632 (50 μM) or thiazovivin (1 μM) were infected with CSFV (MOI = 1). At 24 hpi, cells were fixed and stained with rabbit anti-VIM antibody (red) and mouse anti-dsRNA antibody (green) for confocal microscopy. Bars = 10 μm. The data are presented as the mean ± SD from three independent assays. **, P < 0.01; ***, P < 0.001.