Journal: Cell Reports

Article Title: CDK activity at the centrosome regulates the cell cycle

doi: 10.1016/j.celrep.2024.114066

Figure Lengend Snippet: Artificially restoring Cdc13-HPM SPB localization promotes mitotic entry (A) Experimental outline. (B) Images of the indicated strains 6 h after release from the G1 arrest (7 h after tetracycline addition). Scale bar, 10 μm. (C) DAPI and calcofluor staining of the DNA and septum, respectively, taken at the indicated time after 1-NmPP1 washout. The displayed pixel range is the same for all images of the same strain but not between strains; pixel range is chosen to best represent DNA and septum staining. Scale bars, 10 μm. (D) Cells were heat-fixed at the indicated time points after 1-NmPP1 washout and stained with DAPI and calcofluor to score for passage through mitosis. “Aberrant” includes multiseptate and cut cells. n ≥ 90 cells per time point. See also Figure S2 .

Article Snippet: 1-NmPP1 , Toronto Research Chemicals , Cat# A603003.

Techniques: Staining

Journal: Cell Reports

Article Title: CDK activity at the centrosome regulates the cell cycle

doi: 10.1016/j.celrep.2024.114066

Figure Lengend Snippet:

Article Snippet: 1-NmPP1 , Toronto Research Chemicals , Cat# A603003.

Techniques: Recombinant, Protease Inhibitor, Peptide Fractionation, Mass Spectrometry, Software

Journal: Molecular Biology of the Cell

Article Title: Mps1 promotes poleward chromosome movements in meiotic prometaphase

doi: 10.1091/mbc.E20-08-0525-T

Figure Lengend Snippet: Mps1 promotes microtubule turnover in meiotic prometaphase. (A) In wild-type cells the shortening kinetochores of actively biorienting chromosomes are predicted to cause a high microtubule turnover. MPS1 mutants exhibit a locked-in-place phenotype that might represent a defect in the depolymerization of kinetochore microtubules. (B) Cells that were unable to form bipolar attachments ( spo11 ), and thus in a prolonged prometaphase-like state, were used to measure microtubule turnover. Half spindles of meiotic cells were pulsed with 405 nm light to photoconvert mEos2-Tub1 (from green to red). Images were acquired every 15 s, and the intensity of the red signal was measured (see Materials and Methods ). Scale bar: 2 μm. (C) Microtubule turnover on metaphase spindles was measured in a diploid strain undergoing either meiosis or mitosis. (D) Microtubule turnover was measured in cells expressing STU2-AID* in the presence or absence of auxin and CuSO 4 (copper was used to induce expression of the P CUP1 -AFB2 F-box protein construct). (E) Microtubule turnover was measured on meiotic metaphase spindles of wild-type or mps1-as1 cells in the presence of the Mps1-as1 inhibitor 1-NMPP1. (F) Microtubule turnover was measured on meiotic metaphase and prometaphase spindles of wild-type cells. (G) Microtubule turnover was measured on prometaphase spindles ( spo11 ) in cells with or without the inactivation of Mps1 by 1-NMPP1. (H) Microtubule turnover was measured on prometaphase spindles ( spo11 ) in wild-type or mps1-R170S cells. All experiments show the averages and SEM of three or more biological replicates with three or more cells per replicate (see ).

Article Snippet: Where indicated, auxin (2 mM; Sigma Aldrich I5148-10G), CuSO 4 (200 μM; Sigma Aldrich 451657-10G), or 1-NMPP1 (5 μM; Calbiochem; 5 mM stock in dimethyl sulfoxide) were added to the medium at the time of prophase exit.

Techniques: Expressing, Construct

Journal: Molecular Biology of the Cell

Article Title: Mps1 promotes poleward chromosome movements in meiotic prometaphase

doi: 10.1091/mbc.E20-08-0525-T

Figure Lengend Snippet: Microtubule turnover measurements.

Article Snippet: Where indicated, auxin (2 mM; Sigma Aldrich I5148-10G), CuSO 4 (200 μM; Sigma Aldrich 451657-10G), or 1-NMPP1 (5 μM; Calbiochem; 5 mM stock in dimethyl sulfoxide) were added to the medium at the time of prophase exit.

Techniques: