Fig. 3.

HCC development in LTsc1KO mice is preceded by liver damage, hepatocyte death, inflammation and regeneration. Analysis of male mice at 6 months. (A) Representative H&E-stained sections from Tsc1^fl/fl and LTsc1KO livers. (B) Serum concentrations of ALT and AST are presented as the mean ± SEM (n=8–9). *P<0.05 (ALT) and *P<0.005 (AST). (C,D) Immunostaining and quantification of cleaved caspase-3. Data from ten 200X fields per liver are presented as the mean ± SEM (n=3), *P<0.05. (E) A representative LTsc1KO liver stained with H&E or the macrophage marker F4/80. (F) Inflammation grade in Tsc1^fl/fl and LTsc1KO livers. Data are presented as the mean ± SEM (n=10–12), *P<0.5E–04. (G) Representative H&E and anti-PanCK staining of cholangiocytes in Tsc1^fl/fl livers and expanded progenitor cells (arrows) in LTsc1KO livers. (H) Detection and quantification of proliferating hepatocytes by PCNA immunostaining. Data from five 200X fields per liver are presented as the mean ± SEM (n=3), *P<0.05. (I and J) Detection and quantification of hepatocytes with DNA damage by γ-H2AX immunostaining. Data from ten 200X fields per liver are shown as the mean ± SEM (n=3), *P<0.05. Scale bars, A,C,E and H 100 µm (100X); G and I, 50 µm (200X).