DFO increased HIV-1 Tat-, gp120-, and CQ-induced increases in endolysosome fusion with autolysosomes. (A) Representative fluorescence microscopy reconstructed (using Imaris 3D software) images showing autophagosomes (green puncta), endolysosomes (red puncta), endolysosomes fused with autolysosomes (white puncta), and nuclei (blue). Qualitatively, DFO increased autophagosome fusion with endolysosomes induced by CQ (30 µM), Tat (100 nM), and gp120 (1 nM). (B) Quantitatively, DFO increased the percentage of endolysosomes fusing with autophagosomes per cell induced significantly (p < 0.0001) by CQ (30 µM), Tat (100 nM), and gp120 (1 nM). Image analysis and quantification was 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 the control group 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 < 0.0001.