Figure 3. Hsp42 localizes exclusively to peripheral mCherry-VHL inclusions. (A) WT cells expressing mCherry-VHL were grown at 30°C and shifted to 37°C (+MG132). The cellular localizations of mCherry-VHL (green) and Hsp42 (red) were determined at the indicated time points. Hsp42 localization was determined by immunofluorescence using Hsp42-specific antibodies. Nuclei were visualized by coexpressing HTB1-cerulean (blue). Juxtanuclear mCherry-VHL foci that do not colocalize with Hsp42 are highlighted by red circles. (B) hsp42Δ or hsp26Δ cells coexpressing mCherry-VHL and Hsp42-FLAG or Hsp26-FLAG as indicated were grown at 30°C and shifted to 37°C (+MG132) for 180 min or to 45°C (+MG132) for 20 min. mCherry-VHL is depicted in green, and sHSP stainings using FLAG antibodies are shown in red. Nuclei were visualized by DAPI staining (blue). The dashed lines indicate the border of respective yeast cells. Bars, 1 µm.
Figure 3.

Hsp42 localizes exclusively to peripheral mCherry-VHL inclusions. (A) WT cells expressing mCherry-VHL were grown at 30°C and shifted to 37°C (+MG132). The cellular localizations of mCherry-VHL (green) and Hsp42 (red) were determined at the indicated time points. Hsp42 localization was determined by immunofluorescence using Hsp42-specific antibodies. Nuclei were visualized by coexpressing HTB1-cerulean (blue). Juxtanuclear mCherry-VHL foci that do not colocalize with Hsp42 are highlighted by red circles. (B) hsp42Δ or hsp26Δ cells coexpressing mCherry-VHL and Hsp42-FLAG or Hsp26-FLAG as indicated were grown at 30°C and shifted to 37°C (+MG132) for 180 min or to 45°C (+MG132) for 20 min. mCherry-VHL is depicted in green, and sHSP stainings using FLAG antibodies are shown in red. Nuclei were visualized by DAPI staining (blue). The dashed lines indicate the border of respective yeast cells. Bars, 1 µm.

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