Fig. 3

Cocaine treatment downregulates the RISC complex in macrophages. A Western blot analysis showing the expression of Sigma-1 receptor in untreated T-cells, DCs and macrophages. Actin served as 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 Western blot images showing expression of Dicer, TRBP2, GW182 and Ago2 in macrophages and E T-cells with or without HIV-1 infection and cocaine treatment. Actin served as loading control. D, F Quantitative analysis of the Western blots in (C, E) respectively. The band intensity in each lane was determined by ImageJ software. The percent (%) change of each lane was determined by considering untreated as 100%. G Ago2 immunoprecipitation and Western blot of TRBP2 and GW182 with Western blot of Ago2 as control. AbC—Antibody control; TCL—Total Cell lysate. H Quantitative analysis of the Western blots in (G). The band intensity in each lane was determined by ImageJ software. The percent (%) change of each lane was determined by considering untreated as 100%. I Representative confocal microscopic images showing interaction of Ago2 and GW182 in macrophages with or without HIV-1 infection and cocaine treatment after 6 days. 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)