Fig. 6
From: LPA1-mediated inhibition of CXCR4 attenuates CXCL12-induced signaling and cell migration
Effect of LPA1 activation on CXCR4-mediated migration in cell lines endogenously expressing both receptors. A Expression of CXCR4 and LPAR1 in HEK293A, MDA-MB-231, 8505C, and Hs766t cells was measured using RT-qPCR. The relative expression level of each GPCR was normalized to that of GAPDH. Data represent the mean ± SEM of n = 3 independent experiments. B-E The effect of LPA1 stimulation by alkyl-OMPT on CXCL12-induced cell migration was assessed using a transwell migration assay with alkyl-OMPT (1 μM) in the upper chamber and CXCL12 (10 nM) in the lower chamber (Left panels). The effect of CXCR4 stimulation by CXCL12 on alkyl-OMPT-induced cell migration was assessed using a transwell migration assay with CXCL12 (10 nM) in the upper chamber and alkyl-OMPT (1 μM) in the lower chamber (Right panels). Migrated cells were counted from randomly selected images of 10 fields. Data represent the mean ± SD of n = 3 to 5 independent experiments. F, G The effect of alkyl-OMPT (1 μM) in both the upper and lower chambers on CXCL12 (10 nM)-induced cell migration was assessed in MDA-MB-231 (F) and 8505C cells (G). Data represent the mean ± SD of n = 3 independent experiments. Statistical significance was tested using an unpaired two-tailed Student’s t test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant. H, I CXCL12 (10 nM)-induced cell migration was assessed in the presence of alkyl-OMPT (1 μM) with increasing concentrations of AM966 in the upper chamber. Data represent the mean ± SD of n = 3 independent experiments. AO: alkyl-OMPT; CX: CXCL12