Fig. 4

GPR174 contributes to the metastasis of ESCC through the cAMP-PKA-CREB axis. A Nuclear-cytoplasmic extraction assays and Western blot analyses were applied to quantify the expression of PRKACA in both the nuclear and cytoplasmic of the cells. B The impact of a 24h treatment with 10 μM H-89 (dissolved in DMSO) on the migratory capacity of TE-12 cells overexpressing GPR174 was illustrated, along with the corresponding statistical analysis. Scale bars, 200 μm. C Typical images of Western blotting are shown, demonstrating the expression levels of GPR174, phosphorylated CREB (P-CREB), and total CREB (Total-CREB) in ESCC cells overexpressing GPR174 after a 48h exposure to 10 μM H-89 (dissolved in DMSO). D Statistical analyses and typical images of the Transwell migration and invasion assays for ESCC cells, following a 48h treatment with H-89 (dissolved in DMSO), are presented. Scale bars, 100 μm