CXCL12-CXCR4 signaling pathway in ovarian cancer cells.

<p>A) HeyA8-CXCR4-CBRN/Ar-CBC and parental HeyA8 cells were treated with increasing concentrations of CXCL12 for 10 minutes. Western blot of total cell lysates shows phosphorylated and total AKT, respectively. We used GAPDH as a loading control. Graph shows relative band intensities for phosphorylated AKT in each cell line normalized to total AKT and GAPDH. B) HeyA8-CXCR4-CBRN/Ar-CBC cells were treated with 100 ng/ml CXCL12-α for 0, 5, 15, or 30 minutes in the absence or presence of 1 µM AMD3100. Total cell lysates were probed for phosphorylated and total ERK1/2, respectively. Lysates also were analyzed by Western blot for expression of β-arrestin 2-CBC and endogenous β-arrestin 1 and 2. GAPDH is shown as a loading control. C) Flow cytometry of CXCR4 expression in various HeyA8 cell lines used in this study. Dark line and dashed lines in histogram plots denote isotype control and staining with CXCR4 antibody.