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.