Figure 10.

Stimulation of hepatic death signaling through ASK1, TGF-β, p53, and PI3K/PDK1 by adenoviral delivery of MPK38 in mice. At 5–6 days after injection of adenovirus-expressing green fluorescent protein (GFP) (V) (n = 3), wild-type (WT) MPK38 (n = 3), or kinase-dead (K40R) MPK38 (n = 3), mice were sacrificed and primary hepatocytes were prepared from livers as described in “Materials and methods." Immunoblot analyses were performed using the indicated antibodies to examine the effect of adenoviral delivery of MPK38 on hepatic cell death through ASK1/TGF-β/p53/PDK1 signaling pathways (A), as described in Figs. 3–6. The expression levels of GFP and MPK38, as well as the phosphorylation level of STRAP at Ser¹⁸⁸, were also determined by immunoblot analysis using the indicated antibodies (B). The levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured in mice injected with Ad-GFP, Ad-MPK38, or Ad-K40R (C). Results were expressed as mean ${\displaystyle \pm }$ SEM (*P < 0.05, **P < 0.01 vs. control). Nuclei DNA fragmentation was determined by TUNEL staining, as described in “Materials and methods," in mouse liver 5–6 days after injection of Ad-GFP, Ad-MPK38, or Ad-K40R (D). Control, WT mice; V, vector; Ad, adenovirus.