Fig. 2

a–f CsGRN induces biliary damage and inflammatory cell infiltration both in vitro and in vivo. a The location of injected recombinant plasmid in mice was observed through an in vivo imaging system (IVIS). The color scale indicates the fluorescence intensity of the firefly luciferase gene attached to the plasmid. b Messenger RNA (mRNA) expression levels of CsGRN in liver tissue were measured by quantitative polymerase chain reaction (qPCR). Data are shown as a standard boxplot. c The levels of serum indicators for biliary damage, such as alkaline phosphatase (ALP), alanine aminotransferase (ALT), total bilirubin (TBIL), and total bile acid (TBA), in the mice treated by pCDH-luc vector or pCDH-luc-CsGRN were measured by enzyme-linked immunosorbent assay (ELISA). d Hematoxylin and eosin (HE) staining was used to observe the histological changes in the bile ducts of mice treated by pCDH-luc vector or pCDH-luc-CsGRN (left panels). Scale bar 500 μM. Immunohistochemical (IHC) staining was used to detect the expression of MCP-1 (right panels). Scale bar 100 μM. e Western blot was used to detect the expression of MCP-1, interleukin-6 (IL-6), and cyclooxygenase-2 (COX-2) proteins in the liver. f Immunofluorescence (IF) after double staining with cytokeratin 19 (CK19) and MCP-1 in the portal area of CsGRN-treated mice livers. MCP-1 (green), CK19 (red), DAPI (blue). Scale bar 10 μM. * P < 0.05, ** P < 0.01, *** P < 0.001. For other abbreviations, see Fig. 1