Gin4-mediated phosphorylation of Elm1 is required for normal septin morphology in the absence of SHS1. (A) The sensitivity of shs1∆ cells with Elm1 mutants was tested on SC plates. 10-fold serial dilutions of YEF11766 (shs1∆ GFP-ELM1^WTCDC3-mCherry), YEF11767 (shs1∆ GFP-elm1^S519ACDC3-mCherry), YEF11768 (shs1∆ GFP-elm1^S519DCDC3-mCherry), YEF11769 (shs1∆ GFP-elm1^7ACDC3-mCherry), and YEF11770 (shs1∆ GFP-elm1^7DCDC3-mCherry) cultures were spotted on SC plates and incubated at 25°C (left) and 37°C (right) for 3 d. (B) Representative images of the indicated shs1∆ cells with brightfield (top) and maximum-intensity projections of GFP-tagged Elm1 mutants shown in green (middle) and Cdc3-mCherry shown in magenta (bottom). Strains used are listed in A. The images show maximum projections. The gray dashed line indicates the cell periphery. Scale bars = 2 µm. (C) Montages of representative cells of the indicated strains, with Cdc3-mCherry shown in magenta. Strains used are as follows from top to bottom: YEF11691 (GFP-elm1^7ACDC3-mCherry), YEF11766 (shs1∆ GFP-ELM1^WTCDC3-mCherry), YEF11769 (shs1∆ GFP-elm1^7ACDC3-mCherry), YEF11692 (GFP-elm1^7DCDC3-mCherry), and YEF11770 (shs1∆ GFP-elm1^7DCDC3-mCherry). The images show maximum projections. The yellow arrow indicates septins localized in the bud cortex, the yellow arrowhead indicates asymmetric septins at daughter side. The gray dashed line indicates the cell periphery. Scale bars = 2 µm. (D) Quantification of cells with the indicated septin phenotypes in C. n = number of cells analyzed for each strain.