Fig. 6: YAP1 modulates migration/invasion capability through downstream target AREG.

A Heat map of the expression of YAP1 downstream genes from the TCGA RNA sequencing database in noncancerous and tumor tissues derived from CCA patients. B Correlation between YAP1 and AREG mRNA levels in TCGA_CHOL clinical patients (ρ = 0.46, p = 0.0014). C Treatment with anti-AREG antibody suppressed the invasive ability of HuCCT1 cells. Cells treated with anti-AREG antibodies (1 and 10 µg/mL) were seeded into the upper well of the Boyden chamber with Matrigel and subjected to invasion assays for 18 h. The columns represent mean values from three independent experiments. Bars: means ± SD. D AREG recombinant protein treatment increased the invasive ability of SNU-1196 cells. Cells with AREG recombinant protein (10, 30, and 100 ng/mL) were seeded into the upper well of the Boyden chamber with Matrigel and subjected to invasion assays for 18 h. The columns represent mean values from three independent experiments. Bars: means ± SD. E qRT-PCR analysis of AREG mRNA expression in HuCCT1 cells treated with AREG neutralize antibody or regorafenib. F Quantitation of migration/invasion ability of HuCCT1 treated with regorafenib (10 μM) with or without combined AREG recombinant protein (10, 30, and 100 ng/mL). G Calculate the synergy of the combined use of regorafenib and AREG neutralize antibody through the SynergyFinder service. Data are presented as the mean of three independent experiments ± SEM. The significance of the difference was determined using a nonparametric Mann–Whitney U-test. *p < 0.05, **p < 0.01, ***p < 0.001.