To explore the relationship between TWIST1 expression and EMT progression, we performed ICC with ESCC cell lines using antibodies against E-cadherin (Fig. 4A, B) and vimentin (Fig. 4C, D) and quantitated the percentage of E-Cadherin- and vimentin-positive cells (P < 0.01, Fig. 4K). Here, TWIST1 is linked to intrahepatic cholangiocarcinoma.