Impact of BNI5 deletion on the size, structure, and composition of the septin hourglass. Related to Fig. 4. (A) Additional PREM images of WT and bni5∆ cells. (B) Representative images showing localization of different GFP-tagged septin subunits in WT and bni5∆ cells. Strains used: YEF11087 (mRuby2-TUB1 SHS1-GFP), YEF12141 (bni5∆ mRuby2-TUB1 SHS1-GFP), YEF11754 (mScarlet-TUB1 CDC11-GFP), YEF12143 (bni5∆ mScarlet-TUB1 CDC11-GFP), YEF11753 (mScarlet-TUB1 CDC10-GFP), YEF12142 (bni5∆ mScarlet-TUB1 CDC10-GFP), YEF11755 (mScarlet-TUB1 CDC12-GFP), and YEF12144 (bni5∆ mScarlet-TUB1 CDC10-GFP). (C–F) Quantification of fluorescence signal from the indicated GFP-tagged septin subunits: (C) intensity at the bud neck; (D) intensity in the cytoplasm; (E) intensity ratio at the bud neck to the cytoplasm; (F) diameter of the septin hourglass. Strains used are the same as in B. Circles, brown bars, and gray bars represent individual data points, mean values, and SD, respectively. n denotes the number of cells analyzed per strain. P values were determined using a two-sided unpaired t test.