Figure 5

ROCK1 inhibition impairs BPXV-induced cell contraction and viral protein synthesis. (a) Effect of Thiazovivin on MLC2 phosphorylation (activation). Vero cells were either mock-infected or infected with BPXV at MOI of 5. Thiazovivin or vehicle controls were added at 4 hpi. Cell lysates were prepared at 9 hpi and subjected for detection of the p-MLC2 levels in Western blot analysis (ai). The histogram (aii) shows the band intensity of the protein. The blots were quantified by densitometry (ImageJ) and the data are presented as mean with SD. n = 3 independent experiments. See Supplementary Fig. 6e for full blots. (b) Effect of Thiazovivin on BPXV induced cell contraction and levels of viral proteins. HeLa cells were grown in chamber slides and infected with BPXV at an MOI of 5 for 1 h. Thiazovivin was applied at 4 hpi. At 15 hpi, BPXV (FITC) proteins were probed by immunofluorescence assay. Cell morphology and level of viral proteins of Thiazovivin-treated or untreated cells is shown (bi). The histogram shows the relative reduction in cell size (bii) and relative levels of BPXV proteins (biii) in Thiazovivin treated or untreated cells. The area (n = 50 cells) and the intensity of viral proteins (n = 50 cells) were quantified by ImageJ. The data are presented as mean with SD. Pair-wise statistical comparisons were performed using Student's t-test (***P < 0.001; **P < 0.001).