Extended Data Figure 3: Mitochondrial proteases and peptidases are important for efficient dissolution of aggregates after heat shock.
From: Cytosolic proteostasis through importing of misfolded proteins into mitochondria
a, b, Representative images (a) and quantification (b) over time showing that the mitochondrial split-GFP signal of Lsg1–GFP11 increased after 30 min heat shock and gradually diminished during the 90-min recovery after returning to 30 °C. Top, split-GFP images; middle, MTS–mCherry-labelled mitochondria; bottom, merged images. b shows the mean and s.e.m. of the fraction of cells from three experiments that had a split-GFP signal at each time point. A total of 2,153 cells were counted; three biological repeats. c, Representative images (n = 8) of purified aggregates labelled by GFP-tagged FlucSM, bound to purified mitochondria, labelled with MTS–mCherry. d, Quantification of the immunoblot shown in Fig. 3c. e, f, Immunoblots of FlucSM–HA after aggregates purified from Δhsp104 cells were incubated with wild-type mitochondria for various amounts of time as indicated (e) and quantification of the immunoblot (f). g, h, Dissolution curves of aggregates (g) and their half-decay times (h) showing that the deletion of different mitochondrial proteases delayed the dissolution of cytosolic protein aggregates. g shows mean curves; h shows data points and mean half decay times extracted from fitted curves of three biological repeats. Original data for each repeat are available in Supplementary Information. Scale bars, 5 μm. For gel source data, see Supplementary Fig. 1.
