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. 2016 Apr 12;110(7):1605–1614. doi: 10.1016/j.bpj.2016.02.034

Figure 2.

Figure 2

Establishment of unequal distribution of protein aggregates following temperature shift. (A) Sketch illustrating the TS assay and the specific time points at which PA concentration ([Fluo.]) was analyzed (dotted lines): 12 min after temperature shift (indicated by TS) and upon division. (B) Boxplots of PA concentration (measured as [Fluo.]; total fluorescence in foci/cell volume) in mothers (M) and buds (B) at the time points described in (A). The central red line indicates the median; the lower and upper box edges indicate the 25th and 75th percentiles, respectively; the notches indicate 5% confidence intervals; and the whiskers indicate the most extreme data points not considered outliers. (C) Boxplots of the ratio of [Fluo.] in bud versus mother at the same time points (see B for explanation). (D) Typical phase contrast and fluorescence images (green: Hsp104-GFP; red: Cdc10-mCherry) of small-budded (upper) and large-budded (lower) cells at the time points indicated in (A). Scale bar, 4 μm. (E) Median [B. Fluo.]/[M. Fluo.] ratio at division as a function of initial bud size at the time of TS. Black dots represent individual cells and red circles with error bars (statistical error on mean) display the same data after binning data by size. Green and blue dots correspond to the individual cells in the upper and lower panels shown in (D), respectively. (F) Time from TS to cell division for individual cells (black dots) and after binning (red symbols). Inset: growth arrest as a function of bud size upon TS. (G–L) Evolution with time of cell size (G and H), fluorescence concentration (I and J), and number of aggregates (foci) per cell (K and L) for the buds (left column) and the corresponding mothers (right column). Each colored line represents a group of individual time traces of mother-bud pairs grouped according to the initial volume of the bud Vi as indicated in the figure. Total number of cells analyzed = 78. Vertical dashed lines indicate the average time of division for each group of cells. To see this figure in color, go online.