Figure 7.

HIV-1 TAT-mediated defective mitophagy increases microglial activation and elevates proinflammatory cytokines. (a and b) Representative western blots showing the increased expression of AIF1, a microglial activation marker in mPMs pretreated with 5mM 3-MA and 100 nM wortmannin (a), or in cells pretreated with 25 μM Mdivi-1 (b) for 1 h following exposure to HIV-1 TAT for 24 h. (c and d) Representative western blots showing increased expression of AIF1 in mPMs transfected with either Becn1 siRNA or scrambled siRNA (c) or with Pink1 siRNA or scrambled siRNA (d) following exposure to HIV-1 TAT for 24 h. ACTB was probed as a protein loading control for all experiments. (e and f) Representative bar graphs showing the mRNA expression profile of proinflammatory cytokines such as Tnf, Il1b, and Il6 using qPCR in mPMs pretreated with 5 mM of 3-MA and 100 nM of wortmannin (e) or pretreated with 25 μM Mdivi-1 (f) for 1 h following exposure with HIV-1 TAT for 24 h. (g and h) Representative bar graphs showing the mRNA expression profile of proinflammatory cytokines such as Tnf, Il1b, and Il6 using qPCR in mPMs transfected with either Becn1 siRNA and scrambled siRNA (g) or transfected with either Pink1 siRNA or scrambled siRNA (h) following exposure with HIV-1 TAT for 24 h. Gapdh was used as an internal control to normalize the gene expression for all experiments. Nonparametric Kruskal-Wallis One-way ANOVA followed by the Dunn post hoc test was used to determine the statistical significance between multiple groups. P < 0.05 vs. control; #, P < 0.05 vs. HIV-1 TAT.