Fig. 2

Effect of M-Sec knockdown on TNT formation in U87 cells. a U87.CD4.CCR5 (upper) and U87.CD4.CXCR4 cells (lower) were transfected with either control siRNA (Cr pool #2) or M-Sec-specific siRNA (pool, #1, #2, #3, or #4), cultured for 2 days, and analyzed for the expression of M-Sec or actin (as a loading control) by western blotting, followed by densitometric analysis. The band density values are represented as percentages relative to those of the cells transfected with control siRNA (mean ± SD, n = 3). WB, western blotting. b U87.CD4.CCR5 (upper) and U87.CD4.CXCR4 cells (lower) were transfected with the indicated siRNA, cultured for 2 days, and analyzed for the percentage of TNT-positive cells in 3 different fields (mean ± SD, n = 3). *p < 0.05. c U87.CD4.CCR5 (upper) and U87.CD4.CXCR4 cells (lower) were transfected with either control (Cr pool #2) or M-Sec-specific siRNA (#4) and cultured for 2 days. Cell survival was assessed by the MTT assay. OD values are represented as percentages relative to those of cells transfected with control siRNA (mean ± SD, n = 3). n.s., not significant. d U87.CD4.CCR5 cells were transfected with either control (Cr pool #2) or M-Sec-specific siRNA (#4) and cultured for 2 days. The cell surface area (left) and cell circularity (right) were determined using ImageJ 1.52n software. Three different fields were selected, and 20 cells were analyzed in each field (for a total 60 in each group). *p < 0.05. e U87.CD4.CCR5 cells were transfected with either control (Cr pool #2) or M-Sec-specific siRNA (#4) and cultured for 2 days. The cells were infected with JRFL (input: 100 ng/ml p24 Gag), cultured for 2 days, and analyzed for TNT number in 3 different fields. The values are represented as percentages relative to those of the control siRNA-transfected cells of 1 dpi (mean ± SD, n = 3). *p < 0.05. dpi days postinfection