Budding pattern, polarity establishment, and bud formation during RTG. (A) Cartoon of mitosis, meiosis, and RTG. (B) Cartoon of bipolar budding pattern in diploid cells with calcofluor (blue) at the division site. (C and D) Time-lapse images of calcofluor-stained cells during RTG and mitosis, respectively. The same cell is shown before (left) and after (right) budding with angle between the bud scar and new bud. (E) Absolute frequency of budding angles in RTG and mitosis (n = 200 cells each). (F) Time of GTP-Cdc42 accumulation and bud formation during RTG and mitosis. For RTG, time was measured from nutrient addition (n = 100 cells). For mitosis, time was measured from anaphase spindle breakdown (n = 100 cells). Differences between RTG and mitosis for time of GTP-Cdc42 accumulation and bud formation are statistically significant (Mann-Whitney test with computation of exact two-tailed p-value; P < 0.0001). (G and H) Time-course images of GTP-Cdc42 accumulation in a WT cell undergoing RTG and mitosis, respectively. Time in minutes is indicated. DIC, differential interference contrast. Bars, 3 µm.