<p>(A) Individual prefoldin subunit mutants expressing Guk1-7-GFP were incubated at 37°C in the presence of CHX for 2 hours. The percentage of cells with puncta was calculated from 30–50 GFP positive cells. Scale bar represents 5μm. (B) Four thermosensitive alleles were expressed as GFP fusion proteins in the six prefoldin deletion strains. Cells were incubated with CHX at 25°C and 37°C for 2 hours and fluorescence intensity was measured by flow cytometry. (C) Guk1-7-GFP was expressed in wild type, <i>gim3∆</i>, <i>tcp1-1</i>, or <i>tcp4-1</i> cells and incubated with CHX for two hours at either 25°C or 37°C before flow cytometry analysis. (D) Wild type, <i>gim3∆</i>, <i>tcp1-1</i>, and <i>tcp4-1</i> cells expressing Guk1-7-GFP were incubated with CHX for 2 hours at 25°C or 37°C prior to fixation and imaging. Scale bar represents 5μm.</p
