Figure 5.

VL9 peptide enriches HLA-E in early endosomes and lysosomes. HeLa cells were transiently transfected with HLA-E_EGFP or cotransfected with HLA-E_EGFP and different peptide minigenes. Cells were fixed, permeabilized, and stained with antibodies against protein markers of the early endosome (EEA1), late endosome (Rab7), lysosome (LAMP1), or recycling endosome (Rab11). Cells were then stained with Alexa647-conjugated secondary antibody. (A–D) Quantification of colocalization of HLA-E with different marker proteins. The PCC values of each cell and the mean values are shown with 30–60 cells per sample. Statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. Asterisks show the statistical significance between indicated groups: ns, not significant; *, P < 0.05; ****, P < 0.0001. (E and F) Representative confocal micrographs of HLA-E colocalizing with early endosome (E) or lysosome (F) under different conditions. Scale bar = 20 μm. Data shown are representative of two independent experiments.