Figure 6Different types of misfolded proteins, but not amyloids, are processed together through the Q-body pathway
(a) WT cells expressing Ubc9ts-GFP and CHFP-VHL was grown at 28°C in galactose medium and imaged at 37°C in glucose medium. 5 min series of images show Ubc9ts-GFP signal in green and CHFP-VHL signal in red (i). Cells expressing Ubc9-GFP and CHFP-VHL were similarly prepared and imaged 15 min after the shift (ii). Scale bar equals 1 μm. (b) 15 sec interval series of an inset from cell represented in a (top panel) and from a cell co-expressing Luc-GFP and Ubc9-CHFP (bottom panel, and ). Scale bars equal 1 μm (c). WT cells expressing Htt-Q97-CHFP (red) and Ubc9ts-GFP (green) were prepared and imaged as in a. Scale bar equals 1 μm. (d) 15 sec interval series of an inset from cell represented in c (bottom panel) and from a cell co-expressing GFP-VHL and Rnq1-CHFP (top panel, and ). Scale bars equal 1 μm. (e) Two-color Z-stack projection of cells expressing both Htt-Q97-GFP (in green) and Rnq1-CHFP (in red). Scale bar equals 1 μm. (f) GFP signal of WT, ssa1Δssa2Δ, hsp42Δ, hsp104Δ cells expressing Ubc9ts-GFP or Luc-GFP at 37°C; or Htt-Q97-GFP or Rnq1-GFP at 28°C. Scale bars equal 1 μm.(g-h) Percentages of cells with puncta from a population analyzed in f. Respective total population sizes for WT, ssa1ssa2, hsp42, and hsp104 cells are [ (g): orange n=(2418, 375, 3013, 591) cells; yellow n=(1035, 472, 442, 681) cells; (h): purple, n=(971, 519, 1047, 617) cells; green n=(1165, 410, 1235, 853) cells] over three independent experiments.(6 fields counted per experiment) (*) p<0.05 (**) p<0.005 compared to WT for the same indicated time.