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. 2012 Apr 26;8(4):e1002670. doi: 10.1371/journal.pgen.1002670

Figure 3. Destabilization of the septin ring rescues the lethality and cytokinesis defects caused by high levels of Dma2.

Figure 3

A: Serial dilutions of stationary phase cultures of strains with the indicated genotypes were spotted on YEPD (glucose) or YEPRG (galactose) plates and incubated at 25°C for 2 days. B–D: Wild type (W303), GAL1-DMA2 (ySP3018) and GAL1-DMA2 cdc12-1 (ySP5244) cells grown in YEPR medium were arrested in G1 by alpha factor and released in YEPRG at 25°C. Alpha factor (10 µg/ml) was re-added at t = 105′ to the aliquot of culture used to make protein extracts (D), to prevent cells from entering a second cell cycle. At the indicated times, samples were taken for FACS analysis of DNA contents (not shown) and for scoring budding, nuclear division, spindle formation and septin ring deposition by in situ immunofluorescence with anti-tubulin and anti-Cdc11 antibodies, respectively (B). Representative micrographs were taken at t = 210′ (C). Scale bars: 5 µm. Protein extracts were analysed by western blot with anti-Cdc5 and anti-Pgk1 (loading control) antibodies (D).