Fig. 6

NR2F1-dependent regulation on CXCL12/CXCR4 axis. (a) CXCL12, CXCR4, and CXCR7 mRNAs were quantified by a real-time PCR analysis in NR2F1 high or low expression SACC cells and the control. The results were normalized to GAPDH mRNA used as an internal control. The results were expressed as the relative mRNA expression level of CXCL12, CXCR4, or CXCR7. Data are the mean values ± SEM of at least three independent experiments. The asterisks indicate significant differences between the control and NR2F1 high or low expression SACC cells. *p < 0.05. (b) ChIP assay showed that the combination capacity of CXCL12, and CXCR4 compound significantly increased in NR2F1-overexpressed SACC cells by ChIP test, **p < 0.01, while the combination capacity of CXCR7 compound had no change in NR2F1 -overexpressed SACC cells. n.s p > 0.05. (c) Would healing assay for migration activity of CXCL12-overexpressing SACC cells in response to NR2F1 knockdown. The data showed that the overexpression of CXCL12 could rescue the migration of SACC cells. Error bars represent the mean ± SD of triplicate experiments. (d) Transwell assay for invasion activity of CXCL12-overexpressing SACC cells in response to NR2F1 knockdown. The data showed that the overexpression of CXCL12 could rescue the invasion of SACC cells. Error bars represent the mean ± SD of triplicate experiments. (e) CCK-8 assay for proliferation activity of CXCL12-overexpressing SACC cells in response to NR2F1 knockdown. Comparison of the value of OD between siRNA NR2F1, rhSDF-1a + siRNA NR2F1 and control group, showed that the overexpression of CXCL12 could inhibit the proliferation of SACC cells. Error bars represent the mean ± SD of triplicate experiments