Fig. 2

Up-regulated EphA1 expression in HCC cells promotes SDF-1 expression and secretion. A-B. Chemokine mRNA expression profile determined with qRT-PCR. The data represent the mean ± SD of three independent experiments. a HLE cells: The result of ephrinA1-Fc-activated HLE cells is normalized to the expression in the IgG-Fc-activated HLE cells for each chemokine. The red bar represents the relative SDF-1 mRNA expression level in the ephrinA1-Fc-activated HLE cells. Asterisks indicate significant differences (***P < 0.001) with respect to the IgG-Fc-activated HLE cells. B. Huh-7 cells with different EphA1 expression levels: The result is normalized to the expression in Huh-7 cells transfected with EphA1 scrRNA for each chemokine. The red bar represents the relative SDF-1 mRNA expression level in Huh-7 cells transfected with EphA1 siRNA. Asterisks indicate significant differences (**P < 0.01) with respect to the Huh-7 cells transfected with EphA1 scrRNA. b SDF-1 protein concentration, evaluated with ELISA, in conditioned medium from HCC cells with different EphA1 levels. The data represent the mean ± SD of three independent experiments. Asterisks indicate significant differences (**P < 0.01). c IF staining of SDF-1 (green) and DAPI (blue) in HCC cells (top panel: HLE cells; bottom panel: Huh-7 cells). Images were obtained using a confocal laser scanning microscope. Magnification: × 400; scale bar, 20 μm