Journal: Hepatology (Baltimore, Md.)

Article Title: Disruption of the Growth Hormone--Signal Transducer and Activator of Transcription 5--Insulinlike Growth Factor 1 Axis Severely Aggravates Liver Fibrosis in a Mouse Model of Cholestasis

doi: 10.1002/hep.23469

Figure Lengend Snippet: Increased hepatocyte apoptosis in Mdr2−/−;STAT5Δhep livers. (A) Quantification of cells positive for Ki67 in immunohistochemistry showed increased hepatocyte proliferation in Mdr2−/−;STAT5Δhep livers. (B) Western blot analysis indicated increased protein levels of cleaved Casp3 in liver lysates of Mdr2−/−;STAT5Δhep mice. (C) Mdr2−/−;STAT5Δhep liver lysates showed a highly significant increase in caspase 3 activity, as determined by Casp3 activity assay (n = 4/genotype). (D) Increased numbers of cleaved Casp3-positive hepatocytes in tissue sections of Mdr2−/−;STAT5Δhep livers (arrows). (E) Effects of BA and IGF-1 on apoptotic activity in primary hepatocytes from control and Stat5Δhep livers. Casp3 activity was significantly increased after DCA and glycochenodeoxycholic acid treatment in Stat5Δhep hepatocytes, but not in control hepatocytes. IGF-1 treatment reversed the BA-induced increase in Casp3 activity in Stat5Δhep hepatocytes. BA, bile acids; DCA, deoxycholic acid; GCDCA, glycochenodeoxycholic acid; HSC70, heat shock complex 70.

Article Snippet: For quantification of apoptosis, liver protein lysates were incubated with a fluorogenic Caspase 3 substrate (Ac-DEVD-AFC; Alexis Biochemicals, San Diego, CA).

Techniques: Immunohistochemistry, Western Blot, Activity Assay