Fig. 6. Concentrations of (A) HSF1, (B) Hsp70 and (C) actin in liver tissue from G. mirabilis. Levels of HSF1 and actin were determined by western blotting. Briefly, liver tissue was homogenized in 50mmol1^-1 Tris-Cl, pH 6.8, 4% sodium dodecyl sulfate (SDS) and 1 mmol1^-1 phenylmethylsulfonylfluoride (PMSF). Homogenates were boiled for 5 min and centrifuged at 12 000g for 10 min before loading on 7.5% polyacrylamide gels. After electrophoresis, proteins were blotted onto nitrocellulose membranes. Blots were incubated in primary antibody (for antibodies used, see Materials and methods) for 1.5 h and secondary HRP-conjugated antibody for 1 h. Bands were visualized by a chemiluminescence reaction and their densities determined by scanning densitometry. Values are means ± S.E.M. (N=5). Within a graph, values labeled with different letters differ significantly from one another (P<0.05: pairwise t-test).