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Localization of Cdc42-mCherry SW in WT and ydj1 mutants at 27 °C. Arrows mark examples of large-budded cells used for Cdc42 quantification in B and C . Scale bar: 3 µm. A. Cdc42-mCherry SW levels in mother (m) and bud (b) compartments of large-budded ydj1 Δ cells, grown at 27 °C. (left plot) Mean fluorescence intensities from 19 representative mother-bud pairs are plotted. ns, p ≥ 0.05, p aired t test. (right plot) The log 2 mother-to-bud ratio of Cdc42-mCherry SW mean intensity in WT and ydj1 mutants ( n = 52 per strain). The dotted line denotes a symmetric distribution of Cdc42 between mother and bud. **** p < 0.0001 by one-way ANOVA. See also . B. Cdc42 levels in WT and ydj1 mutants, grown at 27 °C to mid-log p hase. Mean fluorescence intensities of Cdc42-mCherry SW in whole cells (mother and bud combined) are plotted. n = 57–60 per strain; **** p < 0.0001, un p aired t-tests. Immunoblotting shows Cdc42-mCherry SW in each strain, detected using <t>polyclonal</t> <t>anti-RFP</t> antibodies, and α-tubulin, a loading control. See also . D. Association of Cdc42-mCherry SW with GFP-Ydj1 detected by a visible IP assay (top panel). A control reaction used extracts containing untagged Ydj1 (bottom panel). The data underlying the graphs can be found in .
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Localization of Cdc42-mCherry SW in WT and ydj1 mutants at 27 °C. Arrows mark examples of large-budded cells used for Cdc42 quantification in B and C . Scale bar: 3 µm. A. Cdc42-mCherry SW levels in mother (m) and bud (b) compartments of large-budded ydj1 Δ cells, grown at 27 °C. (left plot) Mean fluorescence intensities from 19 representative mother-bud pairs are plotted. ns, p ≥ 0.05, p aired t test. (right plot) The log 2 mother-to-bud ratio of Cdc42-mCherry SW mean intensity in WT and ydj1 mutants ( n = 52 per strain). The dotted line denotes a symmetric distribution of Cdc42 between mother and bud. **** p < 0.0001 by one-way ANOVA. See also . B. Cdc42 levels in WT and ydj1 mutants, grown at 27 °C to mid-log p hase. Mean fluorescence intensities of Cdc42-mCherry SW in whole cells (mother and bud combined) are plotted. n = 57–60 per strain; **** p < 0.0001, un p aired t-tests. Immunoblotting shows Cdc42-mCherry SW in each strain, detected using <t>polyclonal</t> <t>anti-RFP</t> antibodies, and α-tubulin, a loading control. See also . D. Association of Cdc42-mCherry SW with GFP-Ydj1 detected by a visible IP assay (top panel). A control reaction used extracts containing untagged Ydj1 (bottom panel). The data underlying the graphs can be found in .
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Localization of Cdc42-mCherry SW in WT and ydj1 mutants at 27 °C. Arrows mark examples of large-budded cells used for Cdc42 quantification in B and C . Scale bar: 3 µm. A. Cdc42-mCherry SW levels in mother (m) and bud (b) compartments of large-budded ydj1 Δ cells, grown at 27 °C. (left plot) Mean fluorescence intensities from 19 representative mother-bud pairs are plotted. ns, p ≥ 0.05, p aired t test. (right plot) The log 2 mother-to-bud ratio of Cdc42-mCherry SW mean intensity in WT and ydj1 mutants ( n = 52 per strain). The dotted line denotes a symmetric distribution of Cdc42 between mother and bud. **** p < 0.0001 by one-way ANOVA. See also . B. Cdc42 levels in WT and ydj1 mutants, grown at 27 °C to mid-log p hase. Mean fluorescence intensities of Cdc42-mCherry SW in whole cells (mother and bud combined) are plotted. n = 57–60 per strain; **** p < 0.0001, un p aired t-tests. Immunoblotting shows Cdc42-mCherry SW in each strain, detected using <t>polyclonal</t> <t>anti-RFP</t> antibodies, and α-tubulin, a loading control. See also . D. Association of Cdc42-mCherry SW with GFP-Ydj1 detected by a visible IP assay (top panel). A control reaction used extracts containing untagged Ydj1 (bottom panel). The data underlying the graphs can be found in .
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Localization of Cdc42-mCherry SW in WT and ydj1 mutants at 27 °C. Arrows mark examples of large-budded cells used for Cdc42 quantification in B and C . Scale bar: 3 µm. A. Cdc42-mCherry SW levels in mother (m) and bud (b) compartments of large-budded ydj1 Δ cells, grown at 27 °C. (left plot) Mean fluorescence intensities from 19 representative mother-bud pairs are plotted. ns, p ≥ 0.05, p aired t test. (right plot) The log 2 mother-to-bud ratio of Cdc42-mCherry SW mean intensity in WT and ydj1 mutants ( n = 52 per strain). The dotted line denotes a symmetric distribution of Cdc42 between mother and bud. **** p < 0.0001 by one-way ANOVA. See also . B. Cdc42 levels in WT and ydj1 mutants, grown at 27 °C to mid-log p hase. Mean fluorescence intensities of Cdc42-mCherry SW in whole cells (mother and bud combined) are plotted. n = 57–60 per strain; **** p < 0.0001, un p aired t-tests. Immunoblotting shows Cdc42-mCherry SW in each strain, detected using <t>polyclonal</t> <t>anti-RFP</t> antibodies, and α-tubulin, a loading control. See also . D. Association of Cdc42-mCherry SW with GFP-Ydj1 detected by a visible IP assay (top panel). A control reaction used extracts containing untagged Ydj1 (bottom panel). The data underlying the graphs can be found in .
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Localization of Cdc42-mCherry SW in WT and ydj1 mutants at 27 °C. Arrows mark examples of large-budded cells used for Cdc42 quantification in B and C . Scale bar: 3 µm. A. Cdc42-mCherry SW levels in mother (m) and bud (b) compartments of large-budded ydj1 Δ cells, grown at 27 °C. (left plot) Mean fluorescence intensities from 19 representative mother-bud pairs are plotted. ns, p ≥ 0.05, p aired t test. (right plot) The log 2 mother-to-bud ratio of Cdc42-mCherry SW mean intensity in WT and ydj1 mutants ( n = 52 per strain). The dotted line denotes a symmetric distribution of Cdc42 between mother and bud. **** p < 0.0001 by one-way ANOVA. See also . B. Cdc42 levels in WT and ydj1 mutants, grown at 27 °C to mid-log p hase. Mean fluorescence intensities of Cdc42-mCherry SW in whole cells (mother and bud combined) are plotted. n = 57–60 per strain; **** p < 0.0001, un p aired t-tests. Immunoblotting shows Cdc42-mCherry SW in each strain, detected using polyclonal anti-RFP antibodies, and α-tubulin, a loading control. See also . D. Association of Cdc42-mCherry SW with GFP-Ydj1 detected by a visible IP assay (top panel). A control reaction used extracts containing untagged Ydj1 (bottom panel). The data underlying the graphs can be found in .

Journal: PLOS Biology

Article Title: Cdc42 interacts with chaperone Ydj1 to enhance its stability and partitioning during asymmetric cell division and aging in yeast

doi: 10.1371/journal.pbio.3003306

Figure Lengend Snippet: Localization of Cdc42-mCherry SW in WT and ydj1 mutants at 27 °C. Arrows mark examples of large-budded cells used for Cdc42 quantification in B and C . Scale bar: 3 µm. A. Cdc42-mCherry SW levels in mother (m) and bud (b) compartments of large-budded ydj1 Δ cells, grown at 27 °C. (left plot) Mean fluorescence intensities from 19 representative mother-bud pairs are plotted. ns, p ≥ 0.05, p aired t test. (right plot) The log 2 mother-to-bud ratio of Cdc42-mCherry SW mean intensity in WT and ydj1 mutants ( n = 52 per strain). The dotted line denotes a symmetric distribution of Cdc42 between mother and bud. **** p < 0.0001 by one-way ANOVA. See also . B. Cdc42 levels in WT and ydj1 mutants, grown at 27 °C to mid-log p hase. Mean fluorescence intensities of Cdc42-mCherry SW in whole cells (mother and bud combined) are plotted. n = 57–60 per strain; **** p < 0.0001, un p aired t-tests. Immunoblotting shows Cdc42-mCherry SW in each strain, detected using polyclonal anti-RFP antibodies, and α-tubulin, a loading control. See also . D. Association of Cdc42-mCherry SW with GFP-Ydj1 detected by a visible IP assay (top panel). A control reaction used extracts containing untagged Ydj1 (bottom panel). The data underlying the graphs can be found in .

Article Snippet: Proteins were separated by SDS-PAGE on 12.5% polyacrylamide gels, and western blots were visualized using the LI-COR Odyssey system (LI-COR Biosciences, Lincoln, Nebraska) with the following antibodies: mouse monoclonal anti-Cdc42 antibody (clone 28-10) (EMD Millipore, MABN2485), rabbit polyclonal anti-RFP antibodies (Rockland, 600-401-379), mouse monoclonal anti-GFP antibodies (GFP-ID2-s) (DSHB, University of Iowa), and mouse monoclonal anti-alpha tubulin antibody (clone 12G10) (DSHB, University of Iowa).

Techniques: Fluorescence, Western Blot, Control