Figure 2: Pathogenic AR accumulates in the liver and pituitary gland of heterozygous Hsf-1-knockout AR-97Q mice.

(a) Immunohistochemistry of AR-97Q and Hsf-1-knockout AR-97Q mice using anti-Hsf-1, anti-Hsp72 and anti-polyglutamine (1C2) antibodies (13 weeks old). The Hsf-1-knockout AR-97Q mice showed the nuclear accumulation of pathogenic AR (yellow arrows) and decreased levels of Hsf-1 and Hsp72, an inducible form of Hsp70, in the liver and pituitary gland. (b) Immunoblotting for AR in wild-type, AR-97Q and Hsf-1-knockout AR-97Q mice (13 weeks old). (c) Quantification of immunoblotting revealed that the expression levels of AR monomer were upregulated in the liver of heterozygous Hsf-1-knockout AR-97Q mice. Unpaired t-test, n=3. (d–g) Blood tests revealed that the liver enzyme levels, including aspartate aminotransferase (AST) (d) and alanine aminotransferase (ALT) (e), were elevated in heterozygous Hsf-1-knockout AR-97Q mice compared with AR-97Q mice (13 weeks old). There were no significant differences in the levels of AST (f) and ALT (g) between wild-type and heterozygous Hsf-1-knockout mice (13 weeks old). (h) Haematoxylin–eosin staining of the liver of heterozygous Hsf-1-knockout and genetically unmodified AR-97Q mice (13 weeks old). (i) Quantitative analysis showed that the size of hepatocytes was reduced in heterozygous Hsf-1-knockout AR-97Q mice compared with AR-97Q mice. (j) Depletion of Hsf-1 did not alter the size of hepatocytes in wild-type mice. Unpaired t-test (n=4) (d–g). More than 1,000 cells from three livers were analysed in each group, unpaired t-test. (i,j). Error bars indicate s.e.m. (c–g,i,j). Scale bars, 50 μm (a,h). NS, not significant.