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Fig. 7. JNK inhibitor II reduces acetic-acid-induced cell death and abolishes Bcl-xL phosphorylation in yeast expressing Bcl-xL, but not in yeast co-expressing PKC-
and Bcl-xL. Yeast cells expressing Bcl-xL (YEplac181 + Bcl-xL; YEp51 + Bcl-xL) and co-expressing PKC- and Bcl-xL (PKC- + Bcl-xL) were pre-treated in the presence of solvent (DMSO) or in the presence of 20 µM JNK inhibitor II for 8 hours followed by treatment with 0 and 300 mM acetic acid for 1 hour at 30°C. Percentage of dead cells was determined by c.f.u. counts as in Fig. 1. Data are the mean ± s.e.m. of five to eight independent experiments with six replicates. Significant differences from those obtained with DMSO are indicated by *P<0.05 (unpaired Student's t-test). For western blot analysis, protein extracts were separated by 15% SDS-PAGE. For Bcl-xL detection an anti-Bcl-xL goat polyclonal antibody, followed by horseradish peroxidase-conjugated rabbit anti-goat IgG, were used. For ß-actin detection, used as a loading control, membranes were stripped and then reprobed with an anti-actin rabbit antibody. Immunoblots were developed by enhanced chemiluminescence.
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