Figure 4.
Fir1 carries Skt5 between the SSDR. (A) As in Fig. 1 C but with cells coexpressing Cbk1-GFP together with Myo1-Cherry in the presence (n = 20) or absence (n = 20) of FIR1. (B) As in Fig. 1 C but with cells coexpressing Boi1-GFP with Myo1-Cherry in the presence (n = 20) or absence (n = 25) of FIR1. (C) As in Fig. 1 C but with cells coexpressing Skt5-GFP with Myo1-Cherry in the presence (n = 22) or absence of FIR1 (n = 27), in the absence of SIZ1 (n = 23), or in the absence of SPA2 (n = 21). (D) Images of GFP-Skt5 associated with the Shs1-mCherry–labeled septin ring shortly before (upper left panels from top to bottom: overlay, GFP-, Cherry-channel) and shortly after ring splitting (upper right panels from top to bottom: overlay, GFP, Cherry channel). m and d indicate mother and daughter cell, respectively. Lower panels: Intensity plots in relative fluorescent units of the GFP (green) and Cherry (red) channels along the arrows orthogonal to the septin rings before (left panel) and after (right panel) septin ring splitting. (E) As in Fig. 1 C but with cells coexpressing Chs3-GFP with Myo1-Cherry in wild type (n = 20), in fir1∆ cells (n = 20), or in skt5∆ cells (n = 20). (F) As in Fig. 1 C but with cells co-expressing Chs3-GFP with Shs1-mCherry in wild type (n = 40) or in fir1∆skt5∆ cells (n = 40). The significance of the differences between the profiles in B, C, E, and F are shown in Fig. S4, B–E. (G) Stills of the bud necks of cells corresponding to the intensity profiles shown in C, E, and F. Images were taken every 2 min. Scale bar = 3 µm. (H) Left panel: Mean fluorescence intensities of calcofluor-stained bud necks of wild-type (n = 29), fir1∆ (n = 27), chs2∆ (n = 25), and chs2∆ fir1∆ cells (n = 28). ****P < 0.0001; **P < 0.01; ns = not significant. Right panel: Representative stills of calcofluor-stained cells from each genotype. Error bars, SD. Scale bar = 3 µm.

Fir1 carries Skt5 between the SSDR. (A) As in Fig. 1 C but with cells coexpressing Cbk1-GFP together with Myo1-Cherry in the presence (n = 20) or absence (n = 20) of FIR1. (B) As in Fig. 1 C but with cells coexpressing Boi1-GFP with Myo1-Cherry in the presence (n = 20) or absence (n = 25) of FIR1. (C) As in Fig. 1 C but with cells coexpressing Skt5-GFP with Myo1-Cherry in the presence (n = 22) or absence of FIR1 (n = 27), in the absence of SIZ1 (n = 23), or in the absence of SPA2 (n = 21). (D) Images of GFP-Skt5 associated with the Shs1-mCherry–labeled septin ring shortly before (upper left panels from top to bottom: overlay, GFP-, Cherry-channel) and shortly after ring splitting (upper right panels from top to bottom: overlay, GFP, Cherry channel). m and d indicate mother and daughter cell, respectively. Lower panels: Intensity plots in relative fluorescent units of the GFP (green) and Cherry (red) channels along the arrows orthogonal to the septin rings before (left panel) and after (right panel) septin ring splitting. (E) As in Fig. 1 C but with cells coexpressing Chs3-GFP with Myo1-Cherry in wild type (n = 20), in fir1∆ cells (n = 20), or in skt5∆ cells (n = 20). (F) As in Fig. 1 C but with cells co-expressing Chs3-GFP with Shs1-mCherry in wild type (n = 40) or in fir1∆skt5∆ cells (n = 40). The significance of the differences between the profiles in B, C, E, and F are shown in Fig. S4, B–E. (G) Stills of the bud necks of cells corresponding to the intensity profiles shown in C, E, and F. Images were taken every 2 min. Scale bar = 3 µm. (H) Left panel: Mean fluorescence intensities of calcofluor-stained bud necks of wild-type (n = 29), fir1∆ (n = 27), chs2∆ (n = 25), and chs2∆ fir1∆ cells (n = 28). ****P < 0.0001; **P < 0.01; ns = not significant. Right panel: Representative stills of calcofluor-stained cells from each genotype. Error bars, SD. Scale bar = 3 µm.

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