Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2011 Jul 1;22(13):2185–2197. doi: 10.1091/mbc.E11-02-0127

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2011 Raspelli et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

“ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.

PMC Copyright notice

FIGURE 3: — Effect of the lack of Dma proteins on cell viability, cell morphology, and Swe1 stability in HU-treated cells. (A) Serial dilutions of stationary-phase cultures of the indicated strains were spotted on YEPD plates containing the indicated HU concentrations and incubated for 4 d at 25°C. Colony-forming units were then counted to determine cell viability. Plotted values are the mean values ± SD (bars) from three independent experiments. (B) Exponentially growing cultures of wild-type (W303), dma1Δ dma2Δ (ySP1569), and dma1Δ dma2Δ swe1Δ (ySP3164) cells were transferred to YEPD medium containing 200 mM HU at 25°C. After 7 h, cell samples were taken for cell morphology analysis (DIC) and for in situ immunofluorescence analysis of nuclei, mitotic spindles (MTs), and septin ring deposition by using DAPI staining, anti-tubulin antibodies and anti-Cdc11 antibodies, respectively. Bar, 5 μm. (C–E) Exponentially growing (cyc) cultures of SWE1-3HA (ySP3427) and dma1Δ dma2Δ SWE1-3HA (yRF661) cells were arrested in G1 by α-factor and released from G1 arrest at 25°C in YEPD medium containing 200 mM HU (time 0). At the indicated times, cell samples were taken for FACS analysis of DNA contents (C), for scoring budding and septin ring formation (D), and for determining Swe1 levels by Western blot analysis with anti-HA and anti-Swi6 (loading control) antibodies (E). (F) Protein extracts of dma1Δ dma2Δ SWE1-3HA cells grown for 3 h in HU-containing medium (input) were immunoprecipitated with anti-HA antibodies and the immunoprecipitate was incubated for 30 min at 30°C with 20 units/μl lambda phosphatase (PP) or with reaction buffer only (no PP).