Yeast formins, Bni1 and Bnr1, are ectopically recruited to patches in cap2Δ cells. (A and B) Maximum intensity projections of spinning-disk confocal images of wild-type and cap2Δ cells expressing Bni1–3xmNeon (A) or Bnr1–3xmNeon (B). The majority of Bni1–3xmNeon signal localizes to the bud cortex and the majority of Bnr1–3xmNeon signal localizes to the bud neck with faint Bnr1 signal visible outside of the bud neck (white arrows) in both backgrounds. (C and D) Dynamics of Bni1–3xmNeon (C; cyan), or Bnr1–3xmNeon (D; cyan) at Arc15-RFP^mScarlet (magenta) labeled patches monitored by TIRF microscopy in wild-type (top row) and cap2Δ cells (bottom row). Each image is a single frame from a time-lapse series. Dashed lines indicate cell boundaries. Yellow boxes indicate example patches measured in E and F. (E and F) Quantification of the relative Bni1–3xmNeon (E) or Bnr1–3xmNeon (F), and Arc15-RFP^mScarlet signals over time of the boxed patches in C and D. (G and H) Quantification of the number of patches with ectopic Bni1–3xmNeon signal (G) or Bnr1–3xmNeon signal (H). Patches with a positive correlation between the Arc15-RFP^mScarlet signal and Bni1–3xmNeon, or Bnr1–3xmNeon, signal during the 2 min observation window were scored as having formins colocalizing with the patch (see Fig. S3 and Materials and methods for details). The number of patches scored (n) is shown. Statistical significance is calculated by the Chi-squared test (; *, P ≤ 0.05; ****, P ≤ 0.0001). Scale bar = 2 μm.