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Fluorescently Labelled Aerolysin (FLAER) Labelling of Candida albicans Cells

Authors: Sneh Lata Singh
Sneh Lata SinghAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
Bio-protocol author page: a4573
 and Sneha Sudha Komath
Sneha Sudha KomathAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
For correspondence: sskomath@mail.jnu.ac.in
Bio-protocol author page: a4574
,  date: 6/5/2017, 312 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2303.

[Abstract] In this protocol we describe a nonradiolabelled labelling of GPI anchor in Candida albicans. The method uses a fluorescent probe to bind specifically to GPI anchors so that the level of GPI-anchored proteins at the cell surface can be measured. The labelling does not need permeabilization of cells and ...

In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae

Authors: Alex Hopke
Alex HopkeAffiliation: Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
Bio-protocol author page: a4321
 and Robert T. Wheeler
Robert T. WheelerAffiliation 1: Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
Affiliation 2: Graduate School of Biomedical Sciences and Engineering, University of Maine, Orono, Maine, USA
For correspondence: robert.wheeler1@maine.edu
Bio-protocol author page: a4322
,  date: 4/5/2017, 627 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2213.

[Abstract] In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of ...

Detection of Hog1 Phosphorylation in Candida albicans in Response to an Antifungal Protein

Authors: Brigitte ME Hayes
Brigitte ME HayesAffiliation: La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
Bio-protocol author page: a1668
 and Nicole L van der Weerden
Nicole L van der WeerdenAffiliation: La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
For correspondence: n.vanderweerden@latrobe.edu.au
Bio-protocol author page: a1669
,  date: 9/20/2014, 2623 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1244.

[Abstract] It is becoming increasingly apparent that stress signalling is important for tolerance of fungal species to antifungal chemicals and proteins. The high-osmolarity glycerol (HOG) pathway responds to a number of stressors including osmotic and oxidative stress. This protocol describes a method to detect ...

ChIP-Seq in Candida albicans

Authors: Sadri Znaidi
Sadri ZnaidiAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
For correspondence: sadri.znaidi@pasteur.fr
Bio-protocol author page: a1427
, Caroline Proux
Caroline ProuxAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
Bio-protocol author page: a1428
, Sandra Weber
Sandra WeberAffiliation: Biochemistry Department, Institute for Research in Immunology and Cancer and University of Montreal, Montreal, Canada
Bio-protocol author page: a1429
, Simon Drouin
Simon DrouinAffiliation: Department of Medicine, Clinical Research Institute of Montreal and University of Montreal, Montreal, Canada
Bio-protocol author page: a1430
, François Robert
François RobertAffiliation: Department of Medicine, Clinical Research Institute of Montreal and University of Montreal, Montreal, Canada
Bio-protocol author page: a1431
, Martine Raymond
Martine RaymondAffiliation: Biochemistry Department, Institute for Research in Immunology and Cancer and University of Montreal, Montreal, Canada
Bio-protocol author page: a1432
, Jean-Yves Coppée
Jean-Yves CoppéeAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
Bio-protocol author page: a1433
 and Christophe d’Enfert
Christophe d’EnfertAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
For correspondence: christophe.denfert@pasteur.fr
Bio-protocol author page: a1434
,  date: 6/20/2014, 5664 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1158.

[Abstract] Systems biology approaches can be used to study the regulatory interactions occurring between many components of the biological system at the whole-genome level and decipher the circuitries implicated in the regulation of cellular processes, including those imparting virulence to opportunistic fungi. ...

In vitro Analysis for Macrophage Binding and Pro-inflammatory Responses to Candida albicans

Author: Mohlopheni J. Marakalala
Mohlopheni J. MarakalalaAffiliation: Department of Immunology and Infectious Diseases, Harvard School of Public Health, Harvard University, Boston, USA
For correspondence: mjmaraka@hsph.harvard.edu
Bio-protocol author page: a1356
,  date: 5/5/2014, 2758 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1123.

[Abstract] Macrophage recognition of Candida albicans (C. albicans) is facilitated by pattern recognition receptors that interact with the fungal pathogen associated molecular patterns (PAMPs). Dectin-1 is the major macrophage receptor that is known to recognize fungal Beta-glucans leading to induction of various ...

Immunoblot Analysis of Histone H4 Acetylation and Histone H2A Phosphorylation in Candida albicans

Authors: Michael Tscherner
Michael TschernerAffiliation: Max F. Perutz Laboratories, Campus Vienna Biocenter, Medical University of Vienna, Vienna, Austria
Bio-protocol author page: a929
 and Karl Kuchler
Karl KuchlerAffiliation: Max F. Perutz Laboratories, Campus Vienna Biocenter, Medical University of Vienna, Vienna, Austria
For correspondence: karl.kuchler@meduniwien.ac.at
Bio-protocol author page: a496
,  date: 10/20/2013, 3617 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.943.

[Abstract] Posttranslational modifications of histones are required for different processes including transcription, replication and DNA damage repair. This protocol describes the preparation of a whole-cell extracts for the fungal pathogen Candida albicans. Furthermore, the extract is used to detect lysine acetylation ...

Preparation of Candida albicans Biofilms for Transmission Electron Microscopy

Authors: Heather T. Taff
Heather T. TaffAffiliation: Department of Medicine, University of Wisconsin, Madison, USA
For correspondence: htaff@wisc.edu
Bio-protocol author page: a333
 and David R. Andes
David R. AndesAffiliation: Department of Medicine, University of Wisconsin, Madison, USA
For correspondence: dra@medicine.wisc.edu
Bio-protocol author page: a689
,  date: 7/20/2013, 4153 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.822.

[Abstract] Transmission Electron Microscopy is a form of microscopy that allows for imaging of distinct portions of an individual cell. For Candida albicans biofilms, it is often used to visualize the cell walls of fixed samples of yeast and hyphae. This protocol describes how to grow, harvest, and fix Candida ...

Candida albicans Mitochondrial Protein Import Assay

Authors: Victoria L Hewitt
Victoria L HewittAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
For correspondence: victoria.hewitt@monash.edu
Bio-protocol author page: a235
, Trevor Lithgow
Trevor LithgowAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
Bio-protocol author page: a236
 and Ana Traven
Ana TravenAffiliation: Department of Biochemistry and Molecular Biology, Monash University, Melbourne (Clayton), Australia
For correspondence: ana.traven@monash.edu
Bio-protocol author page: a237
,  date: 2/20/2013, 3911 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.331.

[Abstract] We have established the use of Candida albicans as a new model system to study mitochondrial biogenesis. This dimorphic yeast provides an excellent system to investigate the coordination of mitochondrial biogenesis with other cellular networks including cellular metabolism and the cell cycle. Unlike ...

Preparation of Mitochondria from Candida albicans

Authors: Victoria L Hewitt
Victoria L HewittAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
For correspondence: victoria.hewitt@monash.edu
Bio-protocol author page: a235
, Trevor Lithgow
Trevor LithgowAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
Bio-protocol author page: a236
 and Ana Traven
Ana TravenAffiliation: Department of Biochemistry and Molecular Biology, Monash University, Melbourne (Clayton), Australia
For correspondence: ana.traven@monash.edu
Bio-protocol author page: a237
,  date: 2/20/2013, 4010 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.330.

[Abstract] Based on the methods of (Daum et al., 1982) and (Hewitt et al., 2012), we have established the use of Candida albicans as a new model system to study mitochondrial biogenesis. This dimorphic yeast provides an excellent system to investigate the coordination of mitochondrial biogenesis with other cellular ...
1 

ChIP-Seq in Candida albicans

Authors: Sadri Znaidi
Sadri ZnaidiAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
For correspondence: sadri.znaidi@pasteur.fr
Bio-protocol author page: a1427
, Caroline Proux
Caroline ProuxAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
Bio-protocol author page: a1428
, Sandra Weber
Sandra WeberAffiliation: Biochemistry Department, Institute for Research in Immunology and Cancer and University of Montreal, Montreal, Canada
Bio-protocol author page: a1429
, Simon Drouin
Simon DrouinAffiliation: Department of Medicine, Clinical Research Institute of Montreal and University of Montreal, Montreal, Canada
Bio-protocol author page: a1430
, François Robert
François RobertAffiliation: Department of Medicine, Clinical Research Institute of Montreal and University of Montreal, Montreal, Canada
Bio-protocol author page: a1431
, Martine Raymond
Martine RaymondAffiliation: Biochemistry Department, Institute for Research in Immunology and Cancer and University of Montreal, Montreal, Canada
Bio-protocol author page: a1432
, Jean-Yves Coppée
Jean-Yves CoppéeAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
Bio-protocol author page: a1433
 and Christophe d’Enfert
Christophe d’EnfertAffiliation: Genomes and Genetics Department, Institut Pasteur, Paris, France
For correspondence: christophe.denfert@pasteur.fr
Bio-protocol author page: a1434
,  date: 6/20/2014, 5664 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1158.

[Abstract] Systems biology approaches can be used to study the regulatory interactions occurring between many components of the biological system at the whole-genome level and decipher the circuitries implicated in the regulation of cellular processes, including those imparting virulence to opportunistic fungi. ...

Preparation of Candida albicans Biofilms for Transmission Electron Microscopy

Authors: Heather T. Taff
Heather T. TaffAffiliation: Department of Medicine, University of Wisconsin, Madison, USA
For correspondence: htaff@wisc.edu
Bio-protocol author page: a333
 and David R. Andes
David R. AndesAffiliation: Department of Medicine, University of Wisconsin, Madison, USA
For correspondence: dra@medicine.wisc.edu
Bio-protocol author page: a689
,  date: 7/20/2013, 4153 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.822.

[Abstract] Transmission Electron Microscopy is a form of microscopy that allows for imaging of distinct portions of an individual cell. For Candida albicans biofilms, it is often used to visualize the cell walls of fixed samples of yeast and hyphae. This protocol describes how to grow, harvest, and fix Candida ...

Preparation of Mitochondria from Candida albicans

Authors: Victoria L Hewitt
Victoria L HewittAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
For correspondence: victoria.hewitt@monash.edu
Bio-protocol author page: a235
, Trevor Lithgow
Trevor LithgowAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
Bio-protocol author page: a236
 and Ana Traven
Ana TravenAffiliation: Department of Biochemistry and Molecular Biology, Monash University, Melbourne (Clayton), Australia
For correspondence: ana.traven@monash.edu
Bio-protocol author page: a237
,  date: 2/20/2013, 4010 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.330.

[Abstract] Based on the methods of (Daum et al., 1982) and (Hewitt et al., 2012), we have established the use of Candida albicans as a new model system to study mitochondrial biogenesis. This dimorphic yeast provides an excellent system to investigate the coordination of mitochondrial biogenesis with other cellular ...

Candida albicans Mitochondrial Protein Import Assay

Authors: Victoria L Hewitt
Victoria L HewittAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
For correspondence: victoria.hewitt@monash.edu
Bio-protocol author page: a235
, Trevor Lithgow
Trevor LithgowAffiliation: Department of Biochemistry and Molecular Biology and Department of Microbiology, Monash University, Melbourne (Clayton), Australia
Bio-protocol author page: a236
 and Ana Traven
Ana TravenAffiliation: Department of Biochemistry and Molecular Biology, Monash University, Melbourne (Clayton), Australia
For correspondence: ana.traven@monash.edu
Bio-protocol author page: a237
,  date: 2/20/2013, 3911 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.331.

[Abstract] We have established the use of Candida albicans as a new model system to study mitochondrial biogenesis. This dimorphic yeast provides an excellent system to investigate the coordination of mitochondrial biogenesis with other cellular networks including cellular metabolism and the cell cycle. Unlike ...

Immunoblot Analysis of Histone H4 Acetylation and Histone H2A Phosphorylation in Candida albicans

Authors: Michael Tscherner
Michael TschernerAffiliation: Max F. Perutz Laboratories, Campus Vienna Biocenter, Medical University of Vienna, Vienna, Austria
Bio-protocol author page: a929
 and Karl Kuchler
Karl KuchlerAffiliation: Max F. Perutz Laboratories, Campus Vienna Biocenter, Medical University of Vienna, Vienna, Austria
For correspondence: karl.kuchler@meduniwien.ac.at
Bio-protocol author page: a496
,  date: 10/20/2013, 3617 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.943.

[Abstract] Posttranslational modifications of histones are required for different processes including transcription, replication and DNA damage repair. This protocol describes the preparation of a whole-cell extracts for the fungal pathogen Candida albicans. Furthermore, the extract is used to detect lysine acetylation ...

In vitro Analysis for Macrophage Binding and Pro-inflammatory Responses to Candida albicans

Author: Mohlopheni J. Marakalala
Mohlopheni J. MarakalalaAffiliation: Department of Immunology and Infectious Diseases, Harvard School of Public Health, Harvard University, Boston, USA
For correspondence: mjmaraka@hsph.harvard.edu
Bio-protocol author page: a1356
,  date: 5/5/2014, 2758 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1123.

[Abstract] Macrophage recognition of Candida albicans (C. albicans) is facilitated by pattern recognition receptors that interact with the fungal pathogen associated molecular patterns (PAMPs). Dectin-1 is the major macrophage receptor that is known to recognize fungal Beta-glucans leading to induction of various ...

Detection of Hog1 Phosphorylation in Candida albicans in Response to an Antifungal Protein

Authors: Brigitte ME Hayes
Brigitte ME HayesAffiliation: La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
Bio-protocol author page: a1668
 and Nicole L van der Weerden
Nicole L van der WeerdenAffiliation: La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Australia
For correspondence: n.vanderweerden@latrobe.edu.au
Bio-protocol author page: a1669
,  date: 9/20/2014, 2623 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1244.

[Abstract] It is becoming increasingly apparent that stress signalling is important for tolerance of fungal species to antifungal chemicals and proteins. The high-osmolarity glycerol (HOG) pathway responds to a number of stressors including osmotic and oxidative stress. This protocol describes a method to detect ...

In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae

Authors: Alex Hopke
Alex HopkeAffiliation: Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
Bio-protocol author page: a4321
 and Robert T. Wheeler
Robert T. WheelerAffiliation 1: Molecular & Biomedical Sciences, University of Maine, Orono, Maine, USA
Affiliation 2: Graduate School of Biomedical Sciences and Engineering, University of Maine, Orono, Maine, USA
For correspondence: robert.wheeler1@maine.edu
Bio-protocol author page: a4322
,  date: 4/5/2017, 627 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2213.

[Abstract] In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of ...

Fluorescently Labelled Aerolysin (FLAER) Labelling of Candida albicans Cells

Authors: Sneh Lata Singh
Sneh Lata SinghAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
Bio-protocol author page: a4573
 and Sneha Sudha Komath
Sneha Sudha KomathAffiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
For correspondence: sskomath@mail.jnu.ac.in
Bio-protocol author page: a4574
,  date: 6/5/2017, 312 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2303.

[Abstract] In this protocol we describe a nonradiolabelled labelling of GPI anchor in Candida albicans. The method uses a fluorescent probe to bind specifically to GPI anchors so that the level of GPI-anchored proteins at the cell surface can be measured. The labelling does not need permeabilization of cells and ...
1 
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