Figure 7

Livers of HCV and HBV patients with cirrhosis and HCC exhibit activation of DCLK1-β-catenin signaling with increased migratory cell phenotypes. (a) Immunohistochemical staining of liver tissues (normal, HCV + cirrhosis, and HCV + HCC) was simultaneously carried out with anti-ABC 8E7 mAb and anti-DCLK1 ab109029 antibodies. The co-staining of DCLK1 (brown) and active β-catenin (red) in the same epithelial cells are highlighted in the lower panel. Similar staining for HBV-positive cirrhosis and HCC are shown in Supplementary Fig. S2. (b) Total lysates (40 μg) of liver tissues from HCV-positive patients (n = 6) with cirrhosis (lanes 2–5) and HCC (lanes 6, 7), and normal liver (lane 1) were subjected for Western blot using antibodies against active and total β-catenin, DCLK1 (ab109029), and actin. (c) Quantitation of soluble E-cadherin level in the media supernatants of Huh7-RFP (gray bar) and Huh7-RFP-DCLK1 (hatched bar) using ELISA kit (Thermo Fisher) after 24 h of cell passage. (d) Western blot analysis of the total lysates of liver tissues as described in section (b) using mouse mAb sc-8426 that detects both full-length (120-kDa) and cleaved/soluble ectodomain (80-kDa) of E-cadherin.