Figure 6.
Local exocytosis underlies PM waviness. (A and B) Representative example of WT crossed with fus1Δ cells. The top left LM pictures show Fus1-sfGFP (green) and mCherry-D4H (red) expressed in the WT cell. Bottom left TEM pictures show the same mating pair. The virtual tomogram slices show PM shapes, segmented with Imod. Most fus1Δ cells, whether h+ (A) or h− (B), exhibit sPM. Scale bars are 5 µm in LM, 10 µm in TEM, and 100 nm in tomogram slices. (C) Vesicle density in WT and fus1Δ prefusion cells. n = 40 cells, P < 0.0001, unpaired t test. Magenta dots are h+ cells; green dots are h− cells. (D) Vesicle size distribution from C (n = 566 vesicles for WT, 297 for fus1Δ). Bin width = 5. (E) Myo52-tdTomato average time-lapse projection fluorescence profiles along shmoo tips of h+ and h− fus1Δ cells crossed to unlabeled WT (n = 13 h+ WT, 13 h− WT, 12 h+ fus1Δ, 9 h− fus1Δ). Right: Myo52 mean fluorescence intensity of the 5 maximum pixels from profiles as on the left (n = 51 h+ fus1Δ, 45 h− fus1Δ cells from four experiments; P values are from Mann–Whitney tests). WT values are identical to Fig. 5, E and F. (F) Fim1-sfGFP average time-lapse projection fluorescence profiles along h+ and h− fus1Δ shmoo tips (n = 13 h+ WT, 13 h− WT, 12 h+ fus1Δ, 9 h− fus1Δ). Right: Fim1 mean fluorescence intensity of 5 maximum pixels from each peak of profiles as on the left (n = 28 h+ WT, 29 h− WT, 25 h+ fus1Δ, 21 h− fus1Δ cells from two experiments; P values from Mann–Whitney tests). WT values are identical to Fig. 5, E and F. (G) Mating and fusion efficiencies over time (n ≥ 500 cells of each genotype from three to five experiments; bars show SD; *, P < 0.05; **, P < 0.01; ***, P < 0.005; ****, P < 0.0001 from unpaired t tests).

Local exocytosis underlies PM waviness. (A and B) Representative example of WT crossed with fus1Δ cells. The top left LM pictures show Fus1-sfGFP (green) and mCherry-D4H (red) expressed in the WT cell. Bottom left TEM pictures show the same mating pair. The virtual tomogram slices show PM shapes, segmented with Imod. Most fus1Δ cells, whether h+ (A) or h− (B), exhibit sPM. Scale bars are 5 µm in LM, 10 µm in TEM, and 100 nm in tomogram slices. (C) Vesicle density in WT and fus1Δ prefusion cells. n = 40 cells, P < 0.0001, unpaired t test. Magenta dots are h+ cells; green dots are h− cells. (D) Vesicle size distribution from C (n = 566 vesicles for WT, 297 for fus1Δ). Bin width = 5. (E) Myo52-tdTomato average time-lapse projection fluorescence profiles along shmoo tips of h+ and h− fus1Δ cells crossed to unlabeled WT (n = 13 h+ WT, 13 h− WT, 12 h+ fus1Δ, 9 h− fus1Δ). Right: Myo52 mean fluorescence intensity of the 5 maximum pixels from profiles as on the left (n = 51 h+ fus1Δ, 45 h− fus1Δ cells from four experiments; P values are from Mann–Whitney tests). WT values are identical to Fig. 5, E and F. (F) Fim1-sfGFP average time-lapse projection fluorescence profiles along h+ and h− fus1Δ shmoo tips (n = 13 h+ WT, 13 h− WT, 12 h+ fus1Δ, 9 h− fus1Δ). Right: Fim1 mean fluorescence intensity of 5 maximum pixels from each peak of profiles as on the left (n = 28 h+ WT, 29 h− WT, 25 h+ fus1Δ, 21 h− fus1Δ cells from two experiments; P values from Mann–Whitney tests). WT values are identical to Fig. 5, E and F. (G) Mating and fusion efficiencies over time (n ≥ 500 cells of each genotype from three to five experiments; bars show SD; *, P < 0.05; **, P < 0.01; ***, P < 0.005; ****, P < 0.0001 from unpaired t tests).

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal