Fig. 2. The lack of fibronectin significantly diminishes survival signals for hepatocytes, mediated by Bcl-xL.

(A) Immunostaining for cleaved caspase 3 (in brown) at 36hrs after injury. Sections were counterstained with hematoxylin. CV, central vein. Bar=50μm.

(B) Analysis of cleaved caspase 3-positive cells. Data are means±S.D. (cells/field; field=0.14mm² [n=5 for each group]). *, P<0.05; **, P<0.01.

(C) Western blot analysis of Bcl-xL in control and fibronectin-null livers post injury under reducing conditions. Pooled samples from 3 mouse livers from each strain were used for the analysis. Band intensity was measured by densitometry, and each Bcl-xL intensity was normalized to HSC70 (loading control). Then the intensity at 0hr in control livers was set to 1. Each intensity is shown relative to the control value.

(D) Upper panels: Immunostaining for Bcl-xL (in red) and DAPI (to visualize cell nuclei, in blue) at 36hrs after injury (at lower magnification). CV, central vein. Bar=50μm. Lower panels: Triple immunofluorescence staining for Bcl-xL (in red), hepatocyte marker albumin (in green), and DAPI (in blue) at 36hr post injury (at higher magnification). Arrowheads indicate Bcl-xL and albumin double- positive hepatocytes (orange to yellow color). CV, central vein. Bar=25μm.

(E) Analysis of Bcl-xL-positive hepatocytes. Data are means±S.D. (cells/field; field=0.14mm² [n=5 for each group]). *, P<0.05.