Figure 5.
Mating type–specific PM asymmetries and vesicle distributions. (A and B) Representative examples of WT cell pairs with one mating type labeled showing wPM versus sPM (A), and convex versus concave PM curvature (B). Top left LM images show Fus1 in green and cell identity in red, based on mCherry-D4H expression. Bottom left TEM pictures show the same mating pair. A virtual z-slice of the tomogram shows PM shapes, segmented with Imod. (C) Average time-lapse projection of Fim1-sfGFP (green) and Myo52-tdTomato (red) expressed in both cells. Cytosolic mTag-BFP2 (blue) is expressed only in h+ cells. Paired Fim1 fluorescence intensities at the shmoo tips are shown on the right (n = 31 pairs in two experiments; P < 0.0001; paired t test). Individual data, mean ± SD, are shown. (D) Average time-lapse projection of Fim1-sfGFP (green) and Myo52-tdTomato (red) present only in one cell type (left: h+; right: h−) mated with unlabeled partner. (E) Fluorescence profile of Fim1 and Myo52 along the shmoo tips of cells as in D (n = 13 h+ and 13 h− cells). (F) Fim1 and Myo52 mean fluorescence intensity of the 5 maximum pixels around each peak from profiles as in E (n = 28 h+ and 29 h− cells from two experiments for Fim1 and n = 58 h+ and 52 h− cells from four experiments for Myo52; P values from Mann–Whitney tests). (G) Vesicles paired density and size distribution in prefusion WT h+/h− cell pairs. n = 14 tomograms (264 vesicles for h−; 350 for h+), P = 0.0003; paired t test. (H) Vesicle paired density and size distribution in prefusion WT h90 cells classified according to PM curvature and waviness. n = 20 tomograms (669 vesicles for sPM/convex; 830 for wPM/concave cells), P < 0.0001; paired t test. Scale bars are 5 µm in LM (1 µm for insets), 10 µm in TEM, and 100 nm in tomogram slices.

Mating type–specific PM asymmetries and vesicle distributions. (A and B) Representative examples of WT cell pairs with one mating type labeled showing wPM versus sPM (A), and convex versus concave PM curvature (B). Top left LM images show Fus1 in green and cell identity in red, based on mCherry-D4H expression. Bottom left TEM pictures show the same mating pair. A virtual z-slice of the tomogram shows PM shapes, segmented with Imod. (C) Average time-lapse projection of Fim1-sfGFP (green) and Myo52-tdTomato (red) expressed in both cells. Cytosolic mTag-BFP2 (blue) is expressed only in h+ cells. Paired Fim1 fluorescence intensities at the shmoo tips are shown on the right (n = 31 pairs in two experiments; P < 0.0001; paired t test). Individual data, mean ± SD, are shown. (D) Average time-lapse projection of Fim1-sfGFP (green) and Myo52-tdTomato (red) present only in one cell type (left: h+; right: h−) mated with unlabeled partner. (E) Fluorescence profile of Fim1 and Myo52 along the shmoo tips of cells as in D (n = 13 h+ and 13 h− cells). (F) Fim1 and Myo52 mean fluorescence intensity of the 5 maximum pixels around each peak from profiles as in E (n = 28 h+ and 29 h− cells from two experiments for Fim1 and n = 58 h+ and 52 h− cells from four experiments for Myo52; P values from Mann–Whitney tests). (G) Vesicles paired density and size distribution in prefusion WT h+/h− cell pairs. n = 14 tomograms (264 vesicles for h−; 350 for h+), P = 0.0003; paired t test. (H) Vesicle paired density and size distribution in prefusion WT h90 cells classified according to PM curvature and waviness. n = 20 tomograms (669 vesicles for sPM/convex; 830 for wPM/concave cells), P < 0.0001; paired t test. Scale bars are 5 µm in LM (1 µm for insets), 10 µm in TEM, and 100 nm in tomogram slices.

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