Figure 7. The actin cytoskeleton is required for aggregate compartmentalization. Reduction of mCherry-VHL (green) foci numbers during prolonged protein folding stress requires actin polymerization. Cells expressing mCherry-VHL were grown at 30°C and shifted to 37°C (+MG132). LatA or the same volume of DMSO as a control was added to the cells before a temperature shift. (A) mCherry-VHL localization was monitored in WT cells and cells containing a mutation in actin-1, which renders the actin cytoskeleton resistant to LatA. (B–D) hsp42Δ cells with or without the actin-1 mutation expressing mCherry-VHL (B) and WT cells expressing Hsp104-mCFP (C) or mCitrine-luciferase (D) were treated as described in A. All fluorescent reporter proteins are shown in green, and nuclei were visualized by coexpressing HTB1-cerulean (red). SC, synthetic complete. The dashed lines indicate the border of respective yeast cells. Bars, 1 µm.
Figure 7.

The actin cytoskeleton is required for aggregate compartmentalization. Reduction of mCherry-VHL (green) foci numbers during prolonged protein folding stress requires actin polymerization. Cells expressing mCherry-VHL were grown at 30°C and shifted to 37°C (+MG132). LatA or the same volume of DMSO as a control was added to the cells before a temperature shift. (A) mCherry-VHL localization was monitored in WT cells and cells containing a mutation in actin-1, which renders the actin cytoskeleton resistant to LatA. (B–D) hsp42Δ cells with or without the actin-1 mutation expressing mCherry-VHL (B) and WT cells expressing Hsp104-mCFP (C) or mCitrine-luciferase (D) were treated as described in A. All fluorescent reporter proteins are shown in green, and nuclei were visualized by coexpressing HTB1-cerulean (red). SC, synthetic complete. The dashed lines indicate the border of respective yeast cells. Bars, 1 µm.

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