Cryptococcal Cell Morphology Affects Host Cell Interactions and Pathogenicity
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{"title"=>"Cryptococcal cell morphology affects host cell interactions and pathogenicity", "type"=>"journal", "authors"=>[{"first_name"=>"Laura H.", "last_name"=>"Okagaki", "scopus_author_id"=>"26027095100"}, {"first_name"=>"Anna K.", "last_name"=>"Strain", "scopus_author_id"=>"14064009700"}, {"first_name"=>"Judith N.", "last_name"=>"Nielsen", "scopus_author_id"=>"37022640300"}, {"first_name"=>"Caroline", "last_name"=>"Charlier", "scopus_author_id"=>"7005717214"}, {"first_name"=>"Nicholas J.", "last_name"=>"Baltes", "scopus_author_id"=>"35785763800"}, {"first_name"=>"Fabrice", "last_name"=>"Chrétien", "scopus_author_id"=>"8770664600"}, {"first_name"=>"Joseph H.", "last_name"=>"Heitman", "scopus_author_id"=>"19634810800"}, {"first_name"=>"Françoise", "last_name"=>"Dromer", "scopus_author_id"=>"7005331181"}, {"first_name"=>"Kirsten N.", "last_name"=>"Nielsen", "scopus_author_id"=>"7401572878"}], "year"=>2010, "source"=>"PLoS Pathogens", "identifiers"=>{"isbn"=>"1553-7366", "pmid"=>"20585559", "scopus"=>"2-s2.0-77954677679", "issn"=>"15537366", "pui"=>"359189079", "sgr"=>"77954677679", "doi"=>"10.1371/journal.ppat.1000953"}, "id"=>"1353d152-5ebe-3695-a3e9-cb1383f17f87", "abstract"=>"<title>Author Summary</title> <p><italic>Cryptococcus neoformans</italic> is a common life-threatening opportunistic human fungal pathogen. <italic>C. neoformans</italic> grows as a budding yeast <italic>in vitro</italic> with typical cell sizes ranging from 5 to 10 µm in diameter. Early reports suggested the presence of enlarged cells in human infections yet the identity of these cells and their role in virulence remained uncharacterized. Changes in cellular morphology are also observed in the mouse inhalation model of cryptococcosis. These enlarged “titan” cells accounted for 20% of the cells in the lungs. Titan cell formation was found to be regulated by a G-protein coupled receptor (GPCR) signal transduction pathway associated with pheromone sensing. Analysis of titan cells revealed uninucleate, polyploid cells that reproduced by budding, suggesting alterations in the regulation of cell growth and mitosis. Titan cell formation also affected host-pathogen interactions by reducing phagocytosis by host mononuclear cells and was correlated with reduced dissemination within the host. These results describe a novel mechanism by which <italic>C. neoformans</italic> undergoes morphogenesis to evade host phagocytosis, leading to the hypothesis that titan cell formation allows survival of a subset of the population and plays a unique and specialized role during infection.</p>", "link"=>"http://www.mendeley.com/research/cryptococcal-cell-morphology-affects-host-cell-interactions-pathogenicity", "reader_count"=>91, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>2, "Researcher"=>21, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>26, "Student > Postgraduate"=>1, "Student > Master"=>15, "Student > Bachelor"=>10, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>2, "Researcher"=>21, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>26, "Student > Postgraduate"=>1, "Student > Master"=>15, "Student > Bachelor"=>10, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>6, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>50, "Medicine and Dentistry"=>17, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Computer Science"=>2, "Immunology and Microbiology"=>10}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>17}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>10}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>50}, "Computer Science"=>{"Computer Science"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>6}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>3, "Brazil"=>1, "Chile"=>1, "India"=>1}, "group_count"=>3}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/844569"], "description"=>"<p><b>A</b>) Mice were coinfected with an approximate 1∶1 ratio of <b>a</b>:α intranasally at a final concentration of 5×10<sup>4</sup> cells. Lung sections were stained with periodic acid Schiff (PAS) 14 days (top) or 3 days (bottom) post-infection. White arrow denotes <i>C. neoformans</i> cells >10 µm in diameter. Black arrow denotes cells ≤10 µm in diameter. Top: bar  = 100 µm, bottom: bar  = 10 µm. <b>B</b>) The number of small cells (≤10 µm) and titan cells (>10 µm) were quantified in single and coinfections at 7 days post-infection. >500 cells were counted per treatment per mouse. Error bars indicate SD from 3 mice per treatment. Asterisk indicates p<0.01 in pair-wise comparisons to all other samples. <b>C</b>) Mice were infected by inhalation with 5×10<sup>4</sup> of either <b>a</b> cells labeled with AlexaFluor 488 (green, <b>a</b>G), α cells labeled with AlexaFluor 594 (red, αR), or coinfected with an approximate 1∶1 ratio of each mating type. At 2–3 days post-infection, animals were sacrificed and unstained tissue sections were examined for fluorescence and cell size. >80 cells were analyzed per mouse per treatment. Data are representative of three independent experiments with three mice per treatment. Error bars indicate SD. Asterisk indicates p<0.01 in pair-wise comparisons to all other samples. <b>D</b>) Mice were infected with 5×10<sup>7</sup> cells by inhalation of either <b>a</b>, α, or <i>ste3</i><b><i>a</i></b><i>Δ</i> cells labeled with AlexaFluor 488 or coinfected with one labeled and one unlabeled strain. Cells obtained by bronchoalveolar lavage (BAL) were fixed and examined by microscopy for green fluorescence and cell size. >500 cells were examined per animal. Error bars indicate SD from four mice per treatment. Asterisk indicates p<0.01 in pair-wise comparisons to all other samples; p-values >0.2 were observed for other pair-wise comparisons.</p>", "links"=>[], "tags"=>["cells", "lungs", "coinfected"], "article_id"=>515006, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Titan_cells_in_the_lungs_of_coinfected_mice_/515006", "title"=>"Titan cells in the lungs of coinfected mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:23:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/420636", "https://ndownloader.figshare.com/files/420658", "https://ndownloader.figshare.com/files/420672", "https://ndownloader.figshare.com/files/420703", "https://ndownloader.figshare.com/files/420727"], "description"=>"<div><p><em>Cryptococcus neoformans</em> is a common life-threatening human fungal pathogen. The size of cryptococcal cells is typically 5 to 10 µm. Cell enlargement was observed <em>in vivo</em>, producing cells up to 100 µm. These morphological changes in cell size affected pathogenicity via reducing phagocytosis by host mononuclear cells, increasing resistance to oxidative and nitrosative stress, and correlated with reduced penetration of the central nervous system. Cell enlargement was stimulated by coinfection with strains of opposite mating type, and <em>ste3</em><b><em>a</em></b><em>Δ</em> pheromone receptor mutant strains had reduced cell enlargement. Finally, analysis of DNA content in this novel cell type revealed that these enlarged cells were polyploid, uninucleate, and produced daughter cells <em>in vivo</em>. These results describe a novel mechanism by which <em>C. neoformans</em> evades host phagocytosis to allow survival of a subset of the population at early stages of infection. Thus, morphological changes play unique and specialized roles during infection.</p></div>", "links"=>[], "tags"=>["cryptococcal", "morphology", "affects", "interactions", "pathogenicity"], "article_id"=>143048, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1000953.s001", "https://dx.doi.org/10.1371/journal.ppat.1000953.s002", "https://dx.doi.org/10.1371/journal.ppat.1000953.s003", "https://dx.doi.org/10.1371/journal.ppat.1000953.s004", "https://dx.doi.org/10.1371/journal.ppat.1000953.s005"], "stats"=>{"downloads"=>4, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Cryptococcal_Cell_Morphology_Affects_Host_Cell_Interactions_and_Pathogenicity/143048", "title"=>"Cryptococcal Cell Morphology Affects Host Cell Interactions and Pathogenicity", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2010-06-17 00:50:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/845100"], "description"=>"<p>Mice were coinfected intranasally with 4×10<sup>7</sup> cryptococcal cells. At 3 days post-infection, BALs were performed and cells were sorted by FACS based on size. 2×10<sup>4</sup> titan cells or small cells were resuspended in 100 µL RMPI. Cryptococcal cells received <b>A</b>) no treatment, <b>B</b>) 10 mM NaNO<sub>3</sub>, <b>C</b>) 1 mM TBHP. At 0, 6, 16, or 24 hours, aliquots of each treatment were plated on YPD agar and colony forming units (CFU) were determined. Error bars indicate SD from three replicates.</p>", "links"=>[], "tags"=>["cells", "resistant", "oxidative", "nitrosative"], "article_id"=>515541, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g006", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Titan_cells_are_resistant_to_oxidative_and_nitrosative_stress_/515541", "title"=>"Titan cells are resistant to oxidative and nitrosative stress.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:32:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/845173"], "description"=>"<p>Mice were infected with 5×10<sup>7</sup> cells by inhalation of an approximate 1∶1 ratio of <b>a:</b>α cells. At 3 days post-infection, mice were sacrificed, BALs performed, and the resulting cells were fixed and DAPI stained for nuclear content. <b>A</b>) Titan cell containing a single nucleus. <b>B</b>) Titan cell early bud formation. <b>C</b>) Nuclear transfer from a mother (titan cell) to a daughter cell. <b>D</b>) Titan cell late bud formation. <b>E</b>) Cytokinesis of a daughter cell from a titan cell. Bar  = 10 µm.</p>", "links"=>[], "tags"=>["cells"], "article_id"=>515622, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g007", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Titan_cells_can_undergo_cell_division_/515622", "title"=>"Titan cells can undergo cell division.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:33:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/844684"], "description"=>"<p><i>C. neoformans </i><b>a</b> and α strains were combined with AlexaFluor 488 (green) or AlexaFluor 594 (red), respectively, and incubated for 20 minutes. Cells were washed with PBS to remove excess dye. Mice were inoculated with an approximate 1∶1 ratio of <b>a:</b>α cells at a final concentration of 5×10<sup>7</sup> cells. At 2 days post-infection animals were sacrificed, lungs extracted, fixed in 10% buffered formalin, paraffin-embedded, and 5 µm sections generated. Host tissues are autofluorescent at both wavelengths resulting in a yellow color upon overlay. White arrows denote fluorescent <i>C. neoformans</i> cells. Bar  = 20 µm.</p>", "links"=>[], "tags"=>["labeled", "cells", "lungs"], "article_id"=>515127, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g002", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fluorescently_labeled_a_green_and_945_red_cells_in_the_lungs_during_coinfection_/515127", "title"=>"Fluorescently labeled a (green) and α (red) cells in the lungs during coinfection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:25:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/844991"], "description"=>"<p>Mice were intranasally infected with either <b>a</b>, α, or <i>ste3</i><b><i>a</i></b><i>Δ</i> cells labeled with AlexaFluor 488 (green) or coinfected with one labeled and one unlabeled strain (four mice per treatment). Cells obtained by BAL were fixed, stained with DAPI, and examined by microscopy for green fluorescence (cell type) and cell size. >500 cells were examined per animal. Bar  = 10 µm <b>A</b>) <i>C. neoformans </i><b>a</b> cells (green) ≤10 µm in diameter were visible inside host phagocytes. Host cells were identified by large blue DAPI stained nuclei. <b>B</b>) Several small α (≤10 µm) cells (green) can be seen inside a single host cell. <b>C</b>) Mating type <b>a</b> titan cells (>10 µm) are seen in contact with host phagocytes but are too large to be phagocytosed. <b>D</b>) Cells obtained by bronchoalveolar lavage (BAL) were fixed and examined by microscopy for green fluorescence and percent phagocytosis. >500 cells were examined per animal. Error bars indicate SD from four mice per treatment. Asterisk indicates p<0.09 in pair-wise comparisons to all other samples; p-values >0.4 were observed for other pair-wise comparisons.</p>", "links"=>[], "tags"=>["phagocytosis", "lungs", "infected"], "article_id"=>515438, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g005", "stats"=>{"downloads"=>3, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Titan_cell_formation_and_phagocytosis_in_the_lungs_of_infected_mice_/515438", "title"=>"Titan cell formation and phagocytosis in the lungs of infected mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:30:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/844779"], "description"=>"<p>Mice were coinfected intranasally with an approximate 1∶1 ratio of <b>A</b>) <b>a:</b>αNAT, <b>B</b>) <i>ste3</i><b><i>a</i></b><i>Δ</i>#1:α, or <b>C</b>) <i>ste3</i><b><i>a</i></b><i>Δ</i>#2:α at a final concentration of 5×10<sup>4</sup> cells. The actual proportion of <b>a</b> cells in the infecting inoculum was determined by growth on selective medium and is plotted as a horizontal dashed line. At 21 days post-infection animals were sacrificed, the lungs, brain, and spleen were homogenized and serial dilutions plated. >500 colonies per organ per mouse were isolated and assayed for drug resistance to determine mating type. The proportion of <b>a</b> cells is plotted with open circles denoting values from individual animals and bar height representing the geometric mean. To determine <i>P</i>-values, Wilcoxon rank sum analysis was performed on the measured number of <b>a</b> and α cells compared with the expected number, assuming that both strains remained at the initial inoculum proportions.</p>", "links"=>[], "tags"=>["pheromone", "receptor", "mutant", "strains", "penetrate", "cns"], "article_id"=>515223, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_C_neoformans_pheromone_receptor_mutant_strains_penetrate_the_CNS_during_coinfection_/515223", "title"=>"<i>C. neoformans</i> pheromone receptor mutant strains penetrate the CNS during coinfection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:27:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/844894"], "description"=>"<p><b>a</b> or α were combined with AlexaFluor 350 (blue), AlexaFluor 488 (green) or AlexaFluor 594 (red) and incubated for 20 minutes. Mice were inoculated by tail vein injection with an approximate 1∶1 ratio of <b>a</b>:α at a final concentration of 2×10<sup>7</sup> cells. At 1 day post-infection, animals were sacrificed and 50 µm frozen brain sections were obtained. <b>A</b>) Sections from mice infected with <b>a</b> (green) and α (blue) were immunostained with anti-collagen IV primary antibody (endothelial cell membrane) with a TRITC (red) labeled secondary antibody. Bar  = 20 µm <b>B</b>) Sections from mice infected with <b>a</b> (green) and α (red) were imaged by confocal microscopy and sections were compiled as a projection. Bar size  = 20 µm <b>C</b>) Frozen sections from mice infected with <b>a</b> (red) and α (green) were treated with Hoechst (host cell nuclei), imaged by confocal microscopy, and sections were compiled as a 3D rendering. The U-shaped nuclei are indicative of endothelial cells containing cryptococcal cells. Bar  = 10 µm.</p>", "links"=>[], "tags"=>["cells", "endothelial", "blood-brain"], "article_id"=>515333, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g004", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_KN99a_and_KN99_945_cells_interact_with_endothelial_cells_of_the_blood_brain_barrier_during_coinfection_/515333", "title"=>"KN99a and KN99α cells interact with endothelial cells of the blood-brain barrier during coinfection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:28:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/845333"], "description"=>"<p><b>A</b>) Putative pathways involved in titan cell production. <i>Top:</i> Pheromone signaling may enhance an environmental sensing signal leading to titan cell production. <i>Middle:</i> Pheromone signaling may inhibit a negative regulator of titan cell formation. <i>Bottom:</i> Pheromone signaling may act independently of the environmental sensing and/or other pathways leading to titan cells formation. <b>B</b>) The effect of titan cell formation on pathogenesis. <i>Top:</i> Upon initial infection, titan cell production protects a subset of cells from phagocytosis and killing, to establish the infection. <i>Middle: </i><i>C. neoformans</i> cells disseminate from the lungs either as free cells or inside host phagocytes. <i>Bottom:</i> Phagocytosis is required for efficient penetration of the blood-brain barrier. Cryptococcal cells may enter the brain by transcytosis, or as a “Trojan horse” inside of host phagocytes. Over-production of titan cells reduces blood-brain barrier penetration as a result of decreased phagocytosis.</p>", "links"=>[], "tags"=>["titan", "transduction"], "article_id"=>515780, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g009", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Model_of_titan_cell_signal_transduction_and_pathogenesis_/515780", "title"=>"Model of titan cell signal transduction and pathogenesis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:36:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/845264"], "description"=>"<p>Mice were intranasally infected with 5×10<sup>7</sup> cells with an approximate 1∶1 ratio of <b>a:</b>α cells labeled with AlexaFluor 488 (green). At 3 days post-infection, mice were sacrificed, BALs performed, and the resulting cells were fixed. <b>A</b>) Cells were stained with DAPI to measure nuclear content by flow cytometry. Left panel indicates small (≤10 µm) and titan (>20 µm) cell population gates. Right panel indicates DNA content based on DAPI fluorescence for small (dark gray) and titan (white) populations normalized to cell number (% maximum). Dashed lines indicate predicted 1C, 2C, 4C, and 8C DNA content based on DAPI intensity of the 1C and 2C control cells stained and analyzed in the same experiment. <b>B</b>) Cells were grown <i>in vitro</i> in spent DMEM liquid medium for 7 days at 30°C. Cells were fixed and stained with DAPI. Left panel indicates small (≤10 µm), intermediate (>10, ≤20 µm), and titan (>20 µm) cell population gates. Right panel indicates DNA content based on DAPI fluorescence for small (dark gray), intermediate (light gray), and titan (white) populations normalized to cell number. Dashed lines indicate predicted 1C, 2C, 4C, and 8C DNA content based on DAPI intensity of the 1C and 2C control cells strained and analyzed in the same experiment. <b>C</b>) Fixed BAL samples were sorted into small and titan cell populations by fluorescence activated cell sorting. DNA was purified from the sorted populations, normalized to cell number, and chitin synthase 1 (<i>CHS1</i>) gene copy number was determined by comparison to a log phase control sample with a known ratio of 1C:2C cells with a total gene copy number equivalent to 1.4.</p>", "links"=>[], "tags"=>["cells", "dna"], "article_id"=>515713, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.g008", "stats"=>{"downloads"=>1, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Titan_cells_have_increased_DNA_content_/515713", "title"=>"Titan cells have increased DNA content.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-06-17 01:35:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/845425"], "description"=>"<p>PCR Primers.</p>", "links"=>[], "tags"=>["cell biology/cell signaling", "cell biology/morphogenesis and cell biology", "infectious diseases/fungal infections", "infectious diseases/infectious diseases of the nervous system", "infectious diseases/respiratory infections", "microbiology", "microbiology/cellular microbiology and pathogenesis", "microbiology/medical microbiology", "neurological disorders/infectious diseases of the nervous system"], "article_id"=>515858, "categories"=>["Cell Biology", "Medicine", "Microbiology", "Neuroscience"], "users"=>["Laura H. Okagaki", "Anna K. Strain", "Judith N. Nielsen", "Caroline Charlier", "Nicholas J. Baltes", "Fabrice Chrétien", "Joseph Heitman", "Françoise Dromer", "Kirsten Nielsen"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000953.t001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PCR_Primers_/515858", "title"=>"PCR Primers.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2010-06-17 01:37:38"}

PMC Usage Stats | Further Information

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Relative Metric

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