Rga3 localizes to Cdc42 dynamic patches and promotes their exploratory dynamics. (A) Medial spinning-disk time-lapse confocal images of Rga3-GFP and Myo52-tdTomato in mating mixes during early stages of mating. Time is in minutes. Arrowheads highlight the coincidence of Rga3-GFP and Myo52-tdTomato. (B) Distribution of Scd2-GFP zone dwell time in h⁻ WT and rga3Δ cells treated with 0.02 µg/ml P factor on MSL-N pads. n = 4 experiments with n = 10 cells each. (C) Frequency of wandering behavior for Scd2-GFP patches in h⁺ WT and rga3Δ cells mated with h⁻ WT. n ≥ 69 in each of two experiments. Student’s t test P value is indicated. (D) Time-lapse images of Scd2-GFP in h⁺ rga3Δ cells (dashed purple) mated with h⁻ WT (dashed black) showing two examples of Scd2-GFP patch wandering from the cell tip (top and middle) and one example of Scd2-GFP patch assembly–disassembly behavior before stabilization at the cell side (bottom). Green arrowheads indicate wandering patch movement. Yellow arrowheads highlight assembly–disassembly patch dynamics. To the right of the time lapse are kymographs of the rga3Δ cell cortex along the dashed purple arrow. All micrographs are medial-plane spinning-disk confocal images. Error bars show SD. Time is in minutes from start of imaging. Bars, 3 µm.