Fig. 3

EphA1 increases the binding of SDF-1 to CXCR4 in EPCs. a Cell lysates from EPCs co-cultured with IgG-Fc-activated (Lane 1) or ephrinA1-Fc-activated HLE cells (Lane 2) were used for co-IP using anti-SDF-1 antibodies. WB was performed using antibodies against CXCR4, with β-actin as a control for protein loading. The histogram on the right shows the semi-quantification results of WB. The data represent the mean ± SD of three independent experiments. The results show that the activation of EphA1 leads to an increased pull-down of CXCR4 in EPCs. b IF staining of EPCs after co-culturing with IgG-Fc-activated (top panel) or ephrinA1-Fc-activated HLE cells (bottom panel) using anti-CXCR4 antibody (green fluorescence), anti-SDF-1 antibody (red fluorescence), and DAPI (blue fluorescence). Yellow fluorescence in the merged images indicates the co-expression of SDF-1 and CXCR4 in EPCs after co-culturing with HLE cells activated by IgG-Fc or ephrinA1-Fc. Images were obtained using a confocal laser scanning microscope. Magnification: × 400; scale bar, 10 μm