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. 2015 Nov 9;211(3):653–668. doi: 10.1083/jcb.201504073

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© 2015 Ullal et al.

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Figure 6. — Pom1 regulates Fic1 and Pxl1 binding to Cdc15 in vivo. (A) Fic1-GFP in wildtype, pom1^as1, cdc15^19A, and cdc15^19D backgrounds treated with 3MB-PP1 for 4 h at 25°C. Arrows indicate Fic1-GFP localization at the cell tips. (B) Quantification of Fic1-GFP tip localization as in A (n > 10 cells and tips). (C) Fic1-GFP in pom1^as1 cdc25-22 cells arrested in G2 at 36°C for 4 h and treated with methanol (left) or 3MB-PP1 (right) for 10 min. Arrows indicate medial localization of Fic1-GFP. (D) Pxl1-GFP in wild-type, cdc15^19D, and cdc15^16D cells. (E) meGFP-Cdc15 FRAP halftimes (left) and mobile fractions (right) in wild-type, pxl1Δ< and fic1Δ during interphase, ring formation, and constriction. SPB marker Sid4-mRFP was used to determine cytokinesis stage (n = 15). (F) Quantification of the duration of the cytokinetic ring constriction in wildtype, pxl1Δ< and pom1^as1 pxl1Δ cells (n > 10) treated with 3MB-PP1 for 4h. (G) Cells of indicated genotypes treated with 1 µM 3MB-PP1 for 2 h at 25°C, and stained with calcofluor. Arrowheads indicate cells with tip septa. (H) Quantification of tip septa in septated cells as in E (n > 100). Bars, 5 µm.