Fig. 1

Effect of DDX5 knockdown and rescue on HIV-1 infectivity. a Western blot analysis of DDX5 knockdown and rescue. HIV-1 proviral clone pLAI, and siDDX5A with or without increasing concentrations of siDDX5A rescue expressor were transfected into HeLa cells. b HIV-1 infectivity following DDX5 knockdown and rescue. Cells were transfected with siDDX5A and then a second round of siRNA together with pLAI, with or without increasing concentrations of siDDX5A rescue construct. Supernatant was used to infect TZM-bl cells. Relative infectivity was calculated by dividing individual raw luminescence data with that from siControl treated cells. c Supernatant from b was harvested and CA-p24 quantified by ELISA. d Cell lysates from b were harvested and CA-p24 quantified by ELISA. Each graph is a representative of three independent experiments done in triplicate. e Linear regression of mean virion infectivity from triplicate samples on dose of DDX5A rescue plasmid are shown. f Per virion infectiousness was calculated using the virion infectivity (infectivity divided by supernatant CA-p24) and normalised to siControl treated cells. g Supernatant was harvested 48 h after co-transfection of a constant amount of pLAI with increasing concentrations of Myc-DDX5 and used to infect TZM-bl cells. The graph shown is representative of at least two independent experiments done in duplicate. Data is represented as mean of duplicate samples ± SEM. h Cell lysates from g were harvested and subjected to western blotting. i RT-qPCR analysis of unspliced, partially and fully spliced HIV-1 mRNA from left to right respectively. Cells were sequentially transfected with siControl or siDDX5A and then a second round of siRNA together with pLAI. Data shown is representative of three independent repeats, with triplicate samples for each siRNA. Data has been normalised to siControl. Error bars represent ± SEM. Values are scored as a fold-change relative to that of siControl treated cells. Statistical significance: *p < 0.05. See also Additional file 1: Figure S1