Fig. 1

DDX5 interacts with the viral RNA. A Confocal microscopy analysis of SINV (+) RNA and DDX5 protein localization in mock and infected HCT116 cells at 24 hpi by RNA fluorescence in situ hybridization (FISH) (in magenta) combined with protein immunostaining (in green). DAPI staining (in blue) and merge of the different channels are shown. Scale bar, 10 μm. Representative fluorescence intensity profiles of DDX5 (green curve) and SINV RNA (magenta curve) along the yellow line represented on the merge panel and normalized to the maximum value are shown on the right. B Anti DDX5 western blot on total lysate (INPUT), IgG-IP or DDX5-IP samples in both mock and SINV-GFP infected HCT116 cells. C RT-qPCR on SINV genomic (g) RNA upon DDX5 RIP or IgG RIP. Results are expressed as percentage of Input (total RNA) and represent the mean ± standard deviation (SD) of three biological replicates (n = 3). D Anti-dsRNA dot blot assay on serial dilutions of the total RNA (INPUT) and the undiluted RNA samples from DDX5-RIP or IgG-RIP, in mock and SINV infected conditions. J2 antibody was used to detect dsRNAs. E Confocal co-immunofluorescence analysis was performed in mock and SINV infected HCT116 cells using anti-DDX5 rabbit antibody (in green) or anti-J2 mouse antibody (in magenta). Nuclei were stained with DAPI (in blue). Scale bar 10 μm. Representative fluorescence intensity profiles of DDX5 (green curve) and J2 (magenta curve) along the yellow line represented on the merge panel and normalized to the maximum value are shown on the right. One representative experiment out of three is shown in (A-E)