Localization and turnover kinetics of Elm1 and its role in recruiting Bni5 to the bud neck when Bni5–septin association is disrupted. (A) Elm1-GFP accumulation at the budding site. Top: Montages of Elm1-GFP with respect to Cdc3-mCherry were created from selected frames of time-lapse series taken with 1.5-min intervals. Bottom: Quantification of Elm1-GFP and Cdc3-mCherry signals. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Strain used: YEF10440 (ELM1-GFP CDC3-mCherry). (B) FRAP analysis of Elm1-GFP. Top: Lines, symbols, and error bars represent recovery curves, showing mean values and SD, respectively. n denotes the number of cells analyzed per strain. The reference plots of Bni5-C-GFP and Bni5-N-GFP were modified from Fig. 1 F. Bottom: Montages were created from time-lapse series taken with 10-s intervals. Max and T1/2 indicate the estimated maximum amount of recovery and half-time of recovery. Strain used: YEF8437 (ELM1-GFP mRuby2-TUB1). (C) Time-lapse analysis of Bni5-C-GFP and Bni5-N-GFP accumulation at the budding site in elm1Δ cells. Montages were created as described in A. Strains used: YEF9336 (BNI5-C-GFP CDC3-mCherry), YEF9369 (elm1Δ BNI5-C-GFP CDC3-mCherry), YEF10293 (BNI5-N-GFP CDC3-mCherry), and YEF10315 (elm1Δ BNI5-N-GFP CDC3-mCherry). (D) Quantification of GFP-tagged protein recruitment to the bud neck from cells shown in C. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Asterisk indicates variants of Bni5. (E) Elm1-GFP accumulation at the budding site in bni5Δ cells. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Strains used: YEF9305 (ELM1-GFP CDC3-mCherry) and YEF9313 (ELM1-GFP CDC3-mCherry bni5Δ).
Figure 2.

Localization and turnover kinetics of Elm1 and its role in recruiting Bni5 to the bud neck when Bni5–septin association is disrupted. (A) Elm1-GFP accumulation at the budding site. Top: Montages of Elm1-GFP with respect to Cdc3-mCherry were created from selected frames of time-lapse series taken with 1.5-min intervals. Bottom: Quantification of Elm1-GFP and Cdc3-mCherry signals. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Strain used: YEF10440 (ELM1-GFP CDC3-mCherry). (B) FRAP analysis of Elm1-GFP. Top: Lines, symbols, and error bars represent recovery curves, showing mean values and SD, respectively. n denotes the number of cells analyzed per strain. The reference plots of Bni5-C-GFP and Bni5-N-GFP were modified from Fig. 1 F. Bottom: Montages were created from time-lapse series taken with 10-s intervals. Max and T1/2 indicate the estimated maximum amount of recovery and half-time of recovery. Strain used: YEF8437 (ELM1-GFP mRuby2-TUB1). (C) Time-lapse analysis of Bni5-C-GFP and Bni5-N-GFP accumulation at the budding site in elm1Δ cells. Montages were created as described in A. Strains used: YEF9336 (BNI5-C-GFP CDC3-mCherry), YEF9369 (elm1Δ BNI5-C-GFP CDC3-mCherry), YEF10293 (BNI5-N-GFP CDC3-mCherry), and YEF10315 (elm1Δ BNI5-N-GFP CDC3-mCherry). (D) Quantification of GFP-tagged protein recruitment to the bud neck from cells shown in C. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Asterisk indicates variants of Bni5. (E) Elm1-GFP accumulation at the budding site in bni5Δ cells. n denotes the number of cells analyzed per strain. Bold lines and shaded bands represent the mean and SD, respectively. Strains used: YEF9305 (ELM1-GFP CDC3-mCherry) and YEF9313 (ELM1-GFP CDC3-mCherry bni5Δ).

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