Figure 3. Deficiency in cortistatin increases disease severity
in an experimental model of toxic-induced liver fibrosis. Chronic
hepatic fibrosis was induced in wild-type
(CST+/+ ), partially-deficient
(CST+/- ) or totally-deficient
(CST-/- ) mice for cortistatin by injecting
repetitive low doses of the toxic agent CCl4 as depicted
in the diagram. (A ) Survival was daily monitored. (B )
Fibrosis-induced hepatic damage was
quantified by using histopathological Ishak score and the extension of
fibrosis in hematoxylin/eosin (H&E)- and Sirius red-stained liver
sections (5-15 mice/group; scale bars: 200-µm). Fig. S2B displays images
at higher magnification. (C ) Presence of α-smooth muscle actin
(αSMA)-positive myofibroblasts was determined by immunofluorescence
analysis of liver sections (5-13 mice/group; scale bars: 100-µm.).
(D ) Serum levels of direct bilirubin at day 15 (5 mice/group).
(E-F ) Markers of hepatic fibrosis were determined at the
indicated times by measuring collagen contents in liver protein extracts
(5-13 mice/group) and mRNA expression of connective-tissue growth factor
(CTGF), collagen1-α2 (Col1a2) and TGFβ1 (5-11 mice/group).
*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, and Ishak scores that were analyzed
with Mann-Whitney U-test.