Fig. 9. Mlc1 localisation is cell cycle-dependent in yeast and pseudohyphae. (A) Time-lapse video recorded from a pseudohyphal cell coexpressing Nop1-CFP and Mlc1-YFP. Numbers in each frame indicate time elapsed in minutes since inoculation of unbudded cells into fresh medium. The video may be viewed in supplementary material. (B,C) The axial ratio (length/width) of daughter buds of Mlc1-YFP-expressing cells was quantified in cultures growing either as yeast (B) or pseudohyphae (C) and plotted against bud length. In addition, the presence or absence of Mlc1-YFP fluorescence at the tip was recorded. The switch to isotropic growth occurs at a longer bud length and greater axial ratio in pseudohyphae compared to yeast. Note differences in scales. (D) Mlc1-YFP-expressing cells were induced to form hyphal germ tubes. After 105 minutes, cells were fixed and stained with DAPI and examined with a Delta Vision microscope. Mlc1-YFP fluorescence is visible as a spot at the tip and a ring at the site of septation in the germ tube. Nuclei stained with DAPI are visible as circular areas within the mother cell, or migrating into the germ tube, or dividing across the cytokinetic ring. (E) Hyphal germ tube showing Mlc1-YFP simultaneously at the tip and the septation ring. Scale bar, 5 µm (A,E); 15 µm (D).