Fig. 4

Cocaine enhances release of EVs by downregulating BST-2 in HIV-1 infected cells. A Western blot images showing BST-2 expression in T-cells, DCs and macrophages with or without cocaine treatment for 6 days. GAPDH was used as a loading control. B Quantitative analysis of the Western blots in (A). The band intensity in each lane was determined by ImageJ software. The percent (%) change of each lane was determined by considering untreated as 100%. C DCs were treated with increasing concentrations of IFN-α (0—300 IU/ml) and treated with or without 10 µM cocaine for 6 days were analyzed for BST-2 expression by Western Blotting. GAPDH was used as a loading control. D Quantitative analysis of the Western blots in (C). The band intensity in each lane was determined by ImageJ software. The fold change of each lane was determined by considering IFN-α untreated as reference. E U-937 cells were transduced with non-targeted siRNA or BST-2-siRNA and EV release was measured after 3 days by NanoSight. F Protein extracts from U-937 cells after transduction with NT-siRNA and BST-2-siRNA were analyzed for BST-2 expression by Western Blotting. Data represent the mean ± SD of 3 independent experiments, and p-values were calculated relative to untreated controls (*p ≤ 0.05, **p ≤ 0.01, ***p < 0.001)