Fig. 4. Bgs4p localizes to the growing regions: one or both poles, medial ring and septum. (A) CW staining and GFP-Bgs4p localization throughout the mitotic cell cycle. Early log-phase cells (GFP-bgs4⁺ bgs4), grown on YES liquid medium at 28°C, were visualized for CW staining and GFP fluorescence. CW was added at 50 µg/ml followed by immediate examination of the cells. A total of 436 cells from 41 different images were ordered and analyzed. Cells representative of each cell cycle step were selected and ordered to show a cell cycle progression. (B) Magnification of CW staining and GFP-Bgs4p localization to the medial ring and along the plasma membrane during septum formation. (C) Bgs4p localizes to the medial ring later than Bgs1p. CW staining and GFP-Bgs1p or GFP-Bgs4p localization during mitosis and cytokinesis. GFP-bgs1⁺ bgs1 and GFP-bgs4⁺ bgs4 cells were grown as in A, ethanol-fixed, and analyzed for GFP and CW/Hoechst staining. A total of 132 GFP-Bgs1p- and 76 GFP-Bgs4p-expressing mitotic cells, from 35 and 16 different images respectively, were ordered and analyzed. Cells representative of the anaphase to the cytokinesis process were selected and ordered to show a progression. Bar, 2 µm (A,C); 1 µm (B).