Myo1 targeting during cytokinesis depends on Mlc1 and Iqg1. (A) myo1-TD2 localization depends on Mlc1 and Iqg1. Wild-type (WT) (XDY154), mlc1-11 (XDY173), and iqg1Δ (XDY218) cells carrying plasmid pRS316-MYO1-TD2-GFP were grown in SC-Ura medium at 23°C and then examined for myo1-TD2 localization. (B) Full-length Myo1 localization depends on Mlc1 and Iqg1. Strains YEF6179 (mlc1-11 MYO1-GFP CDC3-RFP; left) and YEF6325 (iqg1Δ CDC3-RFP, pRS316-MYO1-C-GFP; right) were grown in liquid SC-Leu media at 23°C and then imaged by 3D dual-color time-lapse microscopy at 23°C with indicated intervals. Montage images of the GFP, RFP, and merged channels from the representative time-lapse data are shown here. Arrowheads indicate the splitting of septin hourglass into two cortical rings, which coincides with mitotic exit and the onset of cytokinesis. (C) myo1-TD2 localization in cells containing different iqg1 alleles. Plasmid pRS315 derivatives carrying the indicated iqg1 alleles (see Table III) were transformed into strain XDY218. Transformants were grown in SC-Ura-Leu medium at 23°C and then quantified for myo1-TD2 localization. APC, APC/C recognition site; CHD, calponin-homology domain; IQ, IQ motifs; Ras-GAP, RasGAP-related domain; RasGAP-C, RasGAP C terminus–related domain.