Fig. 6

NF90/NF45 impedes binding of Ago2 to VEGF mRNA during hypoxia. A Extracts of HEK293T cells treated with different concentrations of CoCl₂, as indicated, were analyzed by Western blot using antibodies to HIF1a and TBP. B Extracts of HEK293T cells transfected with siRNAs targeting NF90/NF45 or a non-targeting control (Scr) and treated with CoCl₂ were analyzed by Western blot using the indicated antibodies. C Immunoprecipitates obtained using anti-Ago2 or control IgG antibodies from extracts described in (B) were analyzed by Western blot using the indicated antibodies. D Total RNA obtained from HEK293T transfected with siRNAs targeting NF90 and NF45 or a non-targeting control (Scr) and treated with CoCl₂ were analyzed by RT-qPCR. Data represent fold mock (IgG) relative to the control samples (siScr), which was attributed a value of 1 (red line), obtained from 3 independent experiments (*P < 0.05, **P < 0.01, independent Student’s t test) (Additional file 9). E RIP analysis of HEK293T cells transfected with NF90/NF45-targeting siRNAs or a non-targeting control (Scr), as indicated, and treated with CoCl₂. RIPs were performed using anti-Ago2 or a control IgG antibody. Immunoprecipitates were analyzed using RT-qPCR. ND indicates ‘Not Detected’. Data represent mean ± SEM obtained from 3 independent experiments (*P < 0.05, **P < 0.01, ***P < 0.001, NS indicates Not Significant, independent Student’s t test) (Additional file 9)