Figure 3.

Immunoblot analysis of tau phosphorylation and solubility after repeated isoflurane treatment in 8.2-mo-old homozygous JNPL3 male mice. Protein extracts from brain stems of control mice (Ctl; lanes 1 and 2, n=10), and mice sampled at 1 wk after the last of four isoflurane treatments (4×A+1w; lanes 3 and 4, n=8) were extracted according to a modified method of Greenberg and Davies (15). Tau from total (A–F), heat-stable soluble (G), and sarkosyl-insoluble (H) fractions were evaluated by immunoblot analysis with the following antibodies: A) AT8 (pS202/pT205); B) CP13 (pS202); C) MC6 (pS235); D) pS262; E) PHF-1 (pS396/pS404); F–H) tau (phospho-independent). Scatterplots represent quantification of the immunoblot bands displayed above them. Levels of phosphoepitopes (A–E) were normalized to total tau. Graphs display immunoreactivity expressed as percentage of control (100%). Numbers in graphs indicate percentage of the tick line with which they are aligned. Data represented are means ± sd (2 representative values displayed; each lane represents an individual mouse). *P < 0.05, **P < 0.01 vs. Ctl; Mann-Whitney U test.