Fig. S4.

Overexpression or disruption of CynA affects cell polarity. (A) The gene encoding CynA was replaced with a Blasticidin S-resistance cassette by homologous recombination (Left), resulting in several independent cynA^- strains. The mutant genotype was confirmed for each strain by Southern blotting, using a piece of flanking DNA at the 3′ end (3′ arm) as a probe (Right). (B) The morphology of cynA^- cells, expressing either the KF2 vector control or CynA-GFP and imaged for 30 min after either 2 or 3 h of differentiation on nonnutrient agar, was quantified using ImageJ. The data shown here is the same dataset as in Fig. 4C, but presented as histograms normalized to the number of cells in each sample (Left), or as cumulative percentages (Right), to show the distribution of data. Each dataset is comprised of between 1,304 and 1,591 cells from a total of nine movies taken from three separate experiments per sample. Cell morphology was quantified using the frame acquired after 30 min of imaging. The data shown do not include the most highly polarized cells, mostly from the population of CynA-GFP–expressing cells at 3 h of differentiation, because these cells were already aggregating and could not be properly segmented.