Fig. 2. Deficiency in cortistatin exacerbates
cholestatic-induced liver fibrosis. Hepatic fibrosis was induced
in wild-type
(CST+/+ ), partially-deficient
(CST+/- ) or totally-deficient
(CST-/- ) mice for cortistatin by using a model
of chronic cholestatic obstruction by bile duct ligation (BDL).
(A ) Survival was monitored and liver and sera were collected as
depicted in the diagram. (B ) Extension of fibrosis was
quantified in Sirius red-stained liver sections (5-15 mice/group, scale
bars: 200-µm). (C ) Histopathological damage and extension of
necrosis area were determined in hematoxylin/eosin (H&E)-stained liver
sections (5-13 mice/group; scale bars: 100-µm). Fig. S2A shows images at
higher magnification. (D ) Serum levels of direct bilirubin were
measured 10 days after BDL (5 mice/group). (E ) Presence of
myofibroblasts was determined by measuring the area with α-smooth muscle
actin (αSMA)-positive immunofluorescence in liver sections (5-7
mice/group, scale bars: 100-µm). (F-G ) Markers of hepatic
fibrosis were determined at the indicated times by measuring collagen
contents in liver protein extracts and mRNA expression of
connective-tissue growth factor (CTGF), αSMA, collagen1-α2 (Col1a2) and
TGFβ1 (5-8 mice/group). Data are the mean±SEM. *p<0.05,
**p<0.01, ***p<0.001 vs.CST+/+ mice. All panels were analyzed with
unpaired two-tailed Student’s t-test, unless survival that was analyzed
with Kaplan-Meier log-rank test.