(A) FRAP analysis of Kar2-sfGFP in wild type mothers (n = 29 cells) and buds (n = 24 cells) under normal growth condition and (B) in the presence of 0.5 µg/ml of Tm. Representative cells are shown. Rectangles indicate the bleached areas. Image show cell before bleaching (pre-bleach), immediately after bleaching (0 s) and 10 s after the initial bleach (10 s). T1/2 represents the time it took to recover 50% of fluorescence of the reached plateau. Graphs of fluorescence recovery of the corresponding cells are shown. (C) Graph indicates the distribution of t1/2 for individual cells of each mutant tested in the presence (+) or absence (−) of 0.5 µg/ml Tm) n >25 cells. n.s. = not significantly different ***p<0.001 (t test). (D) Z-stack projections of CPY*-GFP and corresponding DIC images in wild type cells are shown. More images are shown in Figure 6. (E) Quantification of the FRAP of CPY*-GFP. (F) Single focal plane of Sec61-GFP and Sec61-2-GFP in wild type or ubc7Δ mutant cells. Note that the gray values have been scaled such as to be able to see both proteins. Otherwise, Sec61-2-GFP values are 5–8-fold lower than wild type Sec61-GFP. (G) Distribution of asymmetry indices of Sec61-GFP and Sec61-2-GFP in wild type or ubc7Δ mutant cells ***p<0.0001 (t test). Average ± SD are indicated (A, B, C, E, G). Scale bars = 2 µm.