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Fig. 4. Effect of overexpressed moesin-GFP proteins and moesin knockdown on HIV-1 infection and entry. (A) Effect of FL-, N- and C-moesin-GFP fusions on HIV-1 infection (MOI, 1) in CEM cells. GFP indicates transfection control for virus infection. As further controls, non-transfected cells were either not infected (non-infected) or infection was blocked with neutralizing anti-CD4 Ab (1 µg/ml) or AZT (5 µM). Data are the means ± s.e.m. of four independent experiments carried out in triplicate. (B) Effect of moesin-GFP fusions on HIV-1 infection of PHA-activated PBL (MOI, 1). GFP indicates transfection control for virus infection (defined as 100% infection). Data are the means ± s.e.m. of three independent experiments carried out in triplicate. (C) Effect of silencing endogenous ezrin and/or moesin expression on HIV-1 viral infection in CEM cells. Data are from three independent experiments carried out in triplicate, presented as means ± s.e.m. Scrambled indicates the control. When indicated, infection was inhibited with neutralizing anti-CD4 mAb. (D) Luciferase-based assay of viral entry by non-replicative HIV-1 particles in CEM T cells (left) or primary T cell blasts (right) overexpressing GFP (control, defined as 100% viral entry) or FL- or N-moesin-GFP fusions as indicated. Data are means ± s.e.m. of three independent experiments carried out in triplicate. (E) β-lactamase-based assay of viral entry by non-replicative HIV-1 particles in CEM T cells silenced for moesin (control, defined as 100% viral entry) with X4-tropic HIV-1 envelope (left panel) or VSV-G envelope (right panel). Data are means ± s.e.m. of three independent experiments carried out in triplicate.
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