Fig 3.

Hsp82 protein levels in wild type and mutant strains. (A) Extracts from wild-type and temperature-sensitive hsp82 strains were compared by immunoblotting using rabbit anti-yeast Hsp90 (Ab-4) serum. The bands were quantified by densitometry and normalized to the strain expressing wild-type Hsp82. (B) YCM282/Hsp82 (HSP82), YCM282/G170D (G170D), and YMCiG170D (iG170D) strains with or without an SBA1 expression vector were compared by immunoblotting using the rabbit anti-yeast Hsp90 (Ab-4) serum. The top panel shows the SYPRO ruby dye–stained SDS-PAGE gel containing 2 μg of resolved protein extract from the wild-type strain (HSP82) and 20 μg from the hsp82G170D strains (G170D and iG170D). The middle panel illustrates the relative immunoreactivity corresponding to the amount of Hsp82 or hsp82G170D protein in extracts shown in the top panel. The bands in the immunoblot (middle panel) were measured by densitometry and normalized to reflect differences in protein loading (bottom panel). Hsp, heat shock protein; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis