Modulation of HIV-1 release by anti-CD36 antibody exposure is not related to CD36 interaction with MOxLDL, TSP-1, or type-I collagen. (A) Immunoblot analysis of the phosphorylation of JNK after different stimuli. HIV-1 NLAD8-infected macrophages starved for 2 h in serum-free medium were treated with anti-CD36 mAb, its isotype control (1 µg/ml), or MOxLDL (at 50 µg/ml) for the indicated period of time. α-Tubulin contents on the same immunoblot are presented for control of loading. The molecular weight corresponding to the closest band of the ladder is indicated. (B) Macrophages infected with HIV-1 NLAD8 for 7 d were washed and pretreated for 30 min with medium supplemented or not with FA6-152 mAb or its isotype control at 2 µg/ml. Then medium with or without MOxLDL was added directly onto the cells to obtain the indicated final concentration of MOxLDL and to keep mAb final concentration at 1 µg/ml. Measure of the p24 Gag released during overnight treatment are presented. (C) Cell viability at the end of the experiment shown in B was measured with the CellTiter-Glo kit. (D) Macrophages at 7 d p.i. starved for 2 h in serum-free medium were treated with the anti-CD36 or with the isotype control mAb in complete medium (control), medium supplemented with lipoprotein deficient serum (LPDS), or serum-free medium (no serum). (E) Quantification of the p24 Gag released from HIV-1–infected macrophages treated overnight with the anti-CD36 antibody (FA6-152 at 2 µg/ml), its isotype control, or thrombospondin-1 (TSP-1) and type-I collagen (COL1) at the indicated concentrations. Representative experiments are shown. Except for D, which was performed twice, results were obtained at least three times with different donors. Data are presented as mean ± SEM of triplicates.