Computational protocol: The Production of Monokaryotic Hyphae by Cryptococcus neoformans Can Be Induced by High Temperature Arrest of the Cell Cycle and Is Independent of Same-Sex Mating

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Protocol publication

[…] Cells were harvested, fixed, and stained with propidium iodide (Sigma-Aldrich, St. Louis, MO), and then sorted as described by Sia et al. . Cell cycle analysis was performed using a BD FACSCalibur flow cytometer (Becton Dickinson Biosciences, Sparks, MD). CellQuest Pro software was used for cell collection, and data analysis was performed using ModFit and FlowJo. G1, S, and G2 phases were identified using the Dean-Jett-Fox mathematical model. G2-arrested cells were identified as described , , , , . This population of cells typically shows a large cell phenotype, an increased proportion of cells with 2C DNA content when compared to controls, and a DAPI staining pattern that shows larger nuclei vs. smaller condensed nuclei of G2 phase cells (see below). Cell sortings were performed on a BD FACSAria III (Beckton Dickinson Biosciences) cell sorter and analyzed using BD FACSDiva 6.1 software. Aliquots of sorted live cells were also used to perform the quantitative monokaryotic fruiting assay. All FACS analysis was performed at the Flow Cytometry Core Laboratory at The University of Texas Health Science Center at San Antonio. […]

Pipeline specifications

Software tools FlowJo, BD FACSDiva
Application Flow cytometry
Organisms Cryptococcus neoformans, Homo sapiens, Saccharomyces cerevisiae
Chemicals Nocodazole