FIGURE 1:

Overexpression of Rtn4a, but not Rtn4b or REEP5, increases cell surface localization of integrin β1 and HLA-A. HeLa cells were transiently transfected with plasmids expressing GFP-NLS as a control, Rtn4a-GFP, GFP-Rtn4b, or mCherry-REEP5. (A) Nonpermeabilized cells were stained for surface-localized integrin β1. Here and in some other figures both GFP and mCherry were pseudocolored green to facilitate comparison of merged images. (B) Nonpermeabilized cells were stained for surface-localized HLA-A. (C) Integrin β1 surface fluorescence staining intensity was quantified for 30–66 transfected and nontransfected cells per condition. (D) HLA-A surface fluorescence staining intensity was quantified for 35–69 transfected and nontransfected cells per condition. All representative images are maximum intensity projections of confocal z-stacks. Error bars represent SD. ****, p ≤ 0.0001; NS, not significant.