DFO blocked HIV-1 Tat-, gp120-, and CQ-induced accumulation of damaged mitochondria inside autophagosomes. (A) Imaris 3D reconstructed fluorescence microscopy images showing autophagosomes (green spherical structures), damaged mitochondria (red) inside autophagosomes (red inside green circle), and nuclei (blue). Qualitatively, DFO blocked CQ- (30 µM), Tat- (100 nM), and gp120- (1 nM) induced increases of mitochondria inside autophagosomes. (B) Quantitatively, DFO blocked CQ- (30 µM), Tat- (100 nM), and gp120- (1 nM) induced increases of mean fluorescence intensity (MFI) of mitochondria inside autophagosomes. Image analysis and quantification were performed from five independent experiments with 30 cells quantified for each experimental condition (n = 150). Error bars represent standard deviation (SD) of five independent experiments. A one-way ANOVA multiple comparisons test was used to compare control and treatment groups. Cells were chosen randomly during image acquisition and analysis from the microscope field of view, and no cells were intentionally excluded. Scale bars are 10 µm for the image and 3 µm for the inset. ** p < 0.01 and **** p < 0.0001.