Figure 3
REEP5 and REEP6 might be unnecessary for membrane expression of CXCR1.
(A) HEK293 cells lacking REEP5 and REEP6 were transfected with a GFP-tagged CXCR1 plasmid. Forty-eight hours post-transfection, the cells were fixed with 4% PFA and examined by confocal microscopy. The scale bar denotes 10 μm. (B) FACS analysis of CXCR1 expression. Membrane expression of HA-tagged CXCR1 in HEK293 cells expressing shRNAs was determined with anti-HA and FITC-conjugated secondary antibodies. Counts indicate relative cell number. NS: no staining. (C) A ligand binding assay using 125I-IL-8 was performed in HEK293 cells expressing CXCR1 in the presence of 200 nM unlabeled IL-8. Bonferroni’s Multiple Comparison Test for Vector versus the other groups revealed p > 0.05. The results are from at least three independent experiments.
