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. 2022 Apr 19;11:e67659. doi: 10.7554/eLife.67659

Figure 6. The intracellular fluorescence intensity of Hsp104-RS2 increased linearly and slowly in cells showing stable adenosine triphosphate (ATP) dynamics.

(A) Formation of Hsp104-RS2 foci in wild-type and snf1∆ adk1∆ cells. The RFP signal (magenta) was imaged in log-phase cells grown at 25°C in medium containing 2% glucose. (B) Fraction of cells showing Hsp104-RS2 foci. The percentages of cells with Hsp104-RS2 foci per one field of view (containing 70–342 cells) were plotted. Bars and error bars indicate the mean ± 1SD. The significance of differences was tested using the unpaired two-tailed Welch’s t-test and indicated by the p value. (C) Time-lapse imaging of QUEEN and Hsp104-RS2 in a wild-type cell. (Top) Images of the Hsp104-RS2 signal (inverted grayscale) in the cell at the indicated time points are shown. (Middle) Kymograph of the images shown in the top panel. (Bottom) QUEEN ratio images of the cell. (D) The mean QUEEN ratio and the mean fluorescence intensities of Hsp104-RS2 inside the cell were plotted over time. The dotted line indicates the best linear regression of Hsp104-RS2 intensities with a correlation coefficient of 0.99. (E) Time-lapse imaging of QUEEN and Hsp104-RS2 in an snf1∆ adk1∆ cell showing stable ATP dynamics. (Top) Images of the Hsp104-RS2 signal (inverted grayscale) in the cell at the indicated time points are shown. (Middle) Kymograph of the images shown in the top panel. (Bottom) QUEEN ratio images. (F) The mean QUEEN ratio and the mean fluorescence intensities of Hsp104-RS2 inside the cell were plotted over time. The dotted line indicates the best linear regression of Hsp104-RS2 intensities with a correlation coefficient of 0.99. Scale bar = 5 µm.

Figure 6—source data 1. Raw data for Figure 6.

Figure 6.

Figure 6—figure supplement 1. Simultaneous imaging of adenosine triphosphate (ATP) and protein aggregation in wild-type cells.

Figure 6—figure supplement 1.

(A) The fluorescence intensities of Hsp104-RS2 foci in wild-type (230 foci) and snf1∆ adk1∆ (330 foci) cells grown at 25°C were plotted. The significance of differences was tested using the unpaired two-tailed Welch’s t-test. (B) Four other examples of the data shown in Figure 6D.
Figure 6—figure supplement 1—source data 1. Raw data for Figure 6—figure supplement 1.
Figure 6—figure supplement 2. Simultaneous imaging of adenosine triphosphate (ATP) and protein aggregation in snf1∆ adk1∆ cells showing stable ATP levels.

Figure 6—figure supplement 2.

(A) Four other examples of the data shown in Figure 6F. (B) Comparison of the root mean square deviation (RMSD) of the intracellular fluorescence intensity of Hsp104-RS2 in wild-type cells and snf1∆ adk1∆ cells that showed stable ATP dynamics. The significance of differences was tested using the unpaired two-tailed Welch’s t-test and indicated by a p value.
Figure 6—figure supplement 2—source data 1. Raw data for Figure 6—figure supplement 2.