Fig. 3. Effect of overexpressed moesin-GFP proteins and moesin knockdown on CD4 and CXCR4 cell-surface levels and F-actin distribution in primary lymphocytes. (A) Western blot analysis of specific silencing of endogenous expression of ezrin (oligo 1E) and/or moesin (oligo 2M) 72 hours after siRNA nucleofection of CEM cells. Silencing of endogenous ezrin and/or moesin expression is quantified as the band intensity ratios to -tubulin. A representative experiment of four is shown. (B) Confocal analysis of HIV-1 Env-induced F-actin redistribution in CEM cells in which endogenous moesin is suppressed by knockdown (siRNA moesin-2M). Scrambled indicates the control. xy mid-sections are shown. Quantification of HIV-1-induced moesin or Actin capping is indicated ± s.d. (C) CD4 and CXCR4 cell-surface expression levels (red histograms) in PHA-activated PBLs overexpressing GFP or FL-, N- or C-moesin-GFP fusion proteins. A representative flow cytometry analysis of three is shown. Blue histograms indicate IgG negative control. (D) Flow cytometry analysis of the effect of specific moesin-silencing on CD4 and CXCR4 cell surface expression in primary lymphocytes. Data are the means ± s.e.m. of four independent experiments carried out in triplicate.