FIGURE 2:

Rtn4 knockdown decreases cell surface localization of integrin β1 and HLA-A. HeLa cells were transiently cotransfected with siRNA against Rtn4 and Block-iT fluorescent control or with Block-iT alone. (A) Cells were immunostained for Rtn4. (B) Rtn4 immunofluorescence intensity was quantified for 30–36 cells per condition. (C) Nonpermeabilized cells were stained for surface-localized integrin β1. (D) Nonpermeabilized cells were stained for surface-localized HLA-A. (E) Integrin β1 surface fluorescence staining intensity was quantified for 39–51 transfected and nontransfected cells per condition. (F) HLA-A surface fluorescence staining intensity was quantified for 34–51 transfected and nontransfected cells per condition. All representative images are maximum intensity projections of confocal z-stacks. Scale bars are 10 µm. Error bars represent SD. ****, p ≤ 0.0001; **, p ≤ 0.01; NS, not significant.