Myo2 focuses the polarisome at the cortex. (A) Distribution of the GFP fusions to Myo2, Pea2, Spa2, and Bud6 in WT and myo2_RD cells. The ratios of the RFIs of mother/daughter of Myo2-GFP (n_WT =75, n_myo2RD =87), GFP-Pea2 (n_WT = 87, n_myo2RD = 118), Spa2-GFP (n_WT = 110, n_myo2RD = 190), and Bud6-GFP (n_WT = 91, n_myo2RD = 80) show significantly smaller values in WT than in myo2_RD cells, indicating a higher concentration of the GFP fusion proteins in the bud of WT cells. (B) FLIP measurement of GFP-Pea2 in WT and myo2_RD cells. Left: Curves are the fitted mean of single measurements of the RFI in WT (n = 12) and myo2_RD (n = 22) cells. Mother cells were continuously photobleached every 5 s, and z-stack images were taken every second. Right: The calculated FLIP half-times of GFP-Pea2 in WT cells (72.0 s) and myo2_RD cells (70.9 s). Error bars are SD. ***, P < 0.001; **, P < 0.01; ns, not significant (one-way ANOVA followed by Tukey’s multiple comparison test).