Fig. 5

YTHDC1 depletion affects Env and Vpu expression in infected cells. HeLa/LAV cells were transfected with control (Ctrl) or 2 distinct siRNAs targeting YHTDC1 (DC1-1 and DC1-2) and 72 h later, cells were harvested and A HIV-1 viral proteins Env, Gag and Vpu, YTHDC1 and GAPDH were analyzed by western blot (*, nonspecific band) and B YTHDC1 and C HIV-1 proteins band intensities were quantified by phosphoimaging and normalized to GAPDH (n = 3). Fold change of protein levels upon DC1 knock down over the Ctrl are plotted. Data are presented as mean ± S.D. (n = 3). P values were calculated using one sample t-test (*p < 0.05, ***p < 0.001). D HEK293T cells were co-transfected with a control plasmid or a plasmid expressing GFP-YTHDC1 and pNL4-3. 48 h. later cells harvested and cell lysates were analyzed by western blotting for HIV-1 viral proteins Env, Gag and Vpu and Nef. E ELISA quantification of CAp24 in cell lysates (CAp24 Intracellular; n = 4) and released into the cell supernatants (CAp24 Extracellular; n = 6). Data are represented as mean ± S.D. P values were calculated using one sample t-test (***p < 0.001, ****p < 0.0001)