Figure 4.

HIV-1 TAT increases autophagosome formation and decreases autophagic flux in mPMs. (a and b) Representative western blots showing the expression of MAP1LC3B-II (a) and SQSTM1 (b) in mPMs exposed to HIV-1 TAT (50 ng/mL) for 24 h followed by treatment with 400 nM BAF, added in the last 4 h of the 24 h treatment period. ACTB was probed as a loading control for all experiments. (c) mPMs transfected with tandem fluorescent-tagged MAP1LC3B plasmid followed by HIV-1 TAT (50 ng/mL) and treated with rapamycin (10 nM) for 24 h. Scale bar: 5 μm. (d and e) Bar graph showing the number of autophagosomes (d) and autolysosomes (e) in mPMs transfected with tandem fluorescent-tagged MAP1LC3B plasmid and treated with HIV-1 TAT and rapamycin for 24 h. The data are presented as mean ± SEM from 6 independent experiments. Nonparametric Kruskal – Wallis One-way ANOVA followed by the Dunn post hoc test was used to determine the statistical significance between multiple groups and the Wilcoxon test was used to compare between 2 groups: *, P < 0.05 vs. control.