Mutant alleles of the MYO2 gene genetically interact with Δypt11. (A) Growth was analyzed for strains carrying a chromosomal myo2::kanMX4 deletion and the MYO2 WT allele on a plasmid with a URA3 marker together with empty vector (−), a myo2(LQ) plasmid, a myo2-fis1(2µ) plasmid, or both the myo2(LQ) plasmid and the myo2-fis1(2µ) plasmid either in a YPT11 WT background (top) or a Δypt11 deletion background (bottom). Cells were grown overnight in synthetic dextrose minimal medium containing uracil to allow for loss of the URA3-based MYO2 plasmid. Then, 10-fold serial dilutions were spotted on synthetic dextrose plates lacking uracil (left) to allow growth of cells containing the MYO2 WT plasmid or on synthetic dextrose plates containing uracil and 5-FOA (right) to select for cells that have lost the MYO2 WT plasmid. SD − Ura plates were incubated for 3 d at 30°C, and SD + Ura/5-FOA plates were incubated for 5 d at 30°C. (B) Cells were grown and analyzed as in Fig. 2 D. Results are from the same series of experiments. (C) Δypt11 cells carrying a chromosomal deletion of the MYO2 gene contained plasmids expressing WT (MYO2), myo2(Q1233R), or myo2(L1301P). Cells expressing GFP-Sft2 were grown in synthetic dextrose minimal medium to the logarithmic growth phase and analyzed by fluorescence microscopy. (left) DIC image of a representative cell. (right) Maximum intensity projection of a z stack consisting of 10 focal planes. (D) Cells expressing ER GFP were analyzed as in C with the exception that fluorescence images represent a single focal plane that is focused on the bud. (B–D) Quantifications are mean values from three independent experiments (n = 100), and error bars indicate standard deviations. Bars, 5 µm.