Fig. 2

Dimerization and GPI anchor are critical for antiviral activity of bBST2A1. (a) Schematic representations of bBST2A1 mutants. (b) Western blotting analysis (anti-HA) of HEK293T cells transfected with plasmids expressing wild-type (WT) and mutant bBST2A1. Samples were untreated or treated with β-mercaptoethanol (β-ME) prior to analysis. Numbers to the left of (b) represent the positions and sizes (in kDa) of molecular weight markers. (C-E) HEK293T cells were transfected with NL4–3.E-U- (and HIV-1 Env) (c), pcPFV (d) or pcBFV (e), together with wild-type (WT) or mutant bBST2A1 plasmids. 48 h post-transfection, viral supernatants were collected and used to infect TZM-bl (c), PFVL (d) and BFVL (e). Luciferase assays were performed 48 h post-infection. HIV-1 virions containing culture supernatants were also subjected to western blotting using p24 antibody (c). Cells were harvested for western blot using indicated antibodies. Data of three independent experiments are summarized in the bar graph. Data are represented as mean ± SEM. Significant differences between the control and bBST2A1s values were determined using the student’s t test. The threshold for significance was set at p < 0.05. *p < 0.05, **p < 0.01, NS: not significant, P > 0.05