Fig. 3: Aggregate dissolution by Hsp104 determines protein aggregate steady-state size.
From: ATP hydrolysis by yeast Hsp104 determines protein aggregate dissolution and size in vivo
a Cellular ATP declines during starvation (black circles), accompanied by the increase in median aggregate diameters (green squares). b During recovery period, the ATP levels are restored (black circles), and median aggregate sizes decrease with similar dynamics (green squares). In a and b, the individual data points are means of three technical replicates for each of the three biological replicates. The dashed line is connecting means of the three biological replicates at each time point. c Representative images of Hsp104-GFP foci at indicated times of assembly and dissolution. This experiment was performed in three technical replicates for each of the three biological replicates. Black bar represents 8 µm. d The percentage of cells displaying at least one Hsp104-GFP aggregate increased during starvation in 0.02% glucose and decreased back to control level during recovery in 2% glucose. e The percentage of cells bearing at least one Hsp104E687Q-GFP aggregate is increasing during starvation in 0.02% glucose, however, did not decrease during recovery in 2% glucose in the Hsp104E687Q strain. In d and e, n = 1200 cells were screened for aggregates starting from three independent exponential yeast cultures for each condition. Bar height represents mean ± SD from three separate cultures, each performed in triplicate. The mean of three technical replicates for each biological replicate is displayed as single data points. f Cellular ATP declines during starvation in 0.02% glucose, followed by the increase in median aggregate diameters in the Hsp104E687Q strain. g During recovery period, the ATP levels are restored; however, the median aggregate diameter does not decrease in the Hsp104 E687Q strain. In f and g, the displayed data points are the mean of three technical replicates for each of three biological replicates. The dashed line is connecting means of the three biological replicates at each time point. h Representative images of Hsp104E687Q-GFP foci at indicated times of assembly and dissolution. This experiment was performed in three technical replicates for each of the three biological replicates. The black bar represents 8 µm.
