Figure 5
From: Inhibition of HIV-1 replication with stable RNAi-mediated knockdown of autophagy factors
Additive HIV-1 inhibition by simultaneous knockdown of Atg5 and Atg16. A. Single and double knockdown cell lines were challenged with HIV-1 and replication was followed over time by measuring CA-p24 in the culture supernatant. B. Relative mRNA expression levels of Atg5 and Atg16 was determined by RT-qPCR. C. The effect of RNAi-mediated knockdown on cell growth was determined by the competitive cell growth assay. The percentage cell growth reduction is plotted. D. Single and double knockdown cells were used in single cycle infections. Cells were analyzed 48 h post infection for the percentage of CA-p24 cells (left panel), MFI (middle panel) and CA-p24 concentration in the culture supernatant (right panel). E. Autophagy function in the single and double knockdown cells was analyzed in nutrient-rich and starvation conditions (first 2 bars for each cell line) and for autophagic flux, where the degradation step of autophagy is inhibited by protease inhibitors (last 2 bars for each cell line). The intensity of the LC3-II signal was corrected for the GAPDH level on Western blot.