Fig. 5

Effect of DDX5 re-expression on SINV in DDX5 KO HCT116 cells. A Western blot on WT, DDX5 KO HCT116 and DDX5 KO HCT116 stably expressing either V5-DDX5 (V5-DDX5) or BFP (BFP). Antibodies directed against DDX5 and GAPDH antibody were used. B Western blot on SINV-GFP infected DDX5 KO HCT116 overexpressing V5-DDX5 or BFP (24 hpi, MOI 0.1). Antibodies directed against DDX5 and capsid were used. Tubulin antibody was used as loading control. C-D RT-qPCR of SINV genomic (g) RNA (C) and SINV sub-genomic (sg) RNA (D) in SINV-GFP infected DDX5 KO HCT116 stably expressing V5-DDX5 or BFP (MOI 0.1, 24 hpi). Results are expressed as relative to GAPDH and normalized over the siCTRL condition. E SINV-GFP viral titers from SINV-GFP infected DDX5 KO HCT116 stably expressing V5-DDX5 or BFP (MOI 0.1, 24 hpi), quantified by plaque assay. Results in (C-D-E) represent the mean ± standard deviation of three biological replicates (n = 3). ns = non significant, C-D one sample t test or E paired Student’s t test. F SINV-GFP infection kinetics in WT, DDX5 KO HCT116 and DDX5 KO HCT116 stably expressing either V5-DDX5 (V5-DDX5) or BFP (BFP). The relative GFP fluorescence area (expressed in percentage) of the infected cells as a function of time was measured after SINV GFP infection at an MOI of 0.1, every 3 h for 72 h with a CellcyteX automated cell counter and analyser. WT HCT116 cells, black; DDX5 KO HCT116 cells, red; V5-DDX5 cells, purple; BFP cells, blue. One representative experiment out of three is shown in (A and B)