Fig. 6

Effect of DDX17 depletion on SINV in WT and DDX5 KO HCT116 cells. A Western blot on SINV-GFP infected HCT116 cells, treated with siCTRL and siDDX17. Antibodies directed against DDX5, DDX17 and capsid were used. Tubulin antibody was used as loading control. B-C RT-qPCR of SINV genomic (g) RNA (B) and SINV sub-genomic (sg) RNA (C) in siRNA treated SINV-GFP infected HCT116 cells (MOI 0.1, 24 hpi). Results are expressed as relative to GAPDH and normalized over the siCTRL condition. D SINV-GFP viral titers from infected HCT116 cells transfected with the indicated siRNAs, quantified by plaque assay. E Western blot on mock SINV-GFP infected WT and DDX5 KO HCT116 cells. Antibodies directed against DDX5, DDX17 and capsid were used. Tubulin antibody was used as loading control. F Western blot of samples from SINV-GFP infected DDX5 KO HCT116 cells upon siCTRL or siDDX17 transfection. Specific primary antibodies against DDX5, DDX17 and viral capsid were used. Tubulin was used as loading control. Position and molecular weight (KDa) of protein size markers are shown on the left. G-H RT-qPCR of SINV genomic (g) RNA (G) and SINV sub-genomic (sg) RNA (H) in SINV-GFP infected DDX5 KO HCT116 cells (MOI 0.1, 24 hpi), upon siRNA transfection. Results are expressed as relative to GAPDH and normalized to the siCTRL condition. I SINV-GFP viral titers from infected DDX5 KO HCT116 cells transfected with the indicated siRNAs, quantified by plaque assay. Results in (B-C-D and G-H-I) represent the mean ± standard deviation of three biological replicates (n = 3). * p ≤ 0.05, **p ≤ 0.01, B-C and G-H one sample t test or D and I paired Student’s t test. One representative experiment out of three is shown in (A, E and F)