title: Spatially transformed fluorescence image data for ERK-MAPK and selected proteins within human epidermis.
abstract: BACKGROUND
  Phosphoprotein signalling pathways have been intensively studied in vitro, yet their role in regulating tissue homeostasis is not fully understood. In the skin, interfollicular keratinocytes differentiate over approximately 2 weeks as they traverse the epidermis. The extracellular signal-regulated kinase (ERK) branch of the mitogen-activated protein kinase (MAPK) pathway has been implicated in this process. Therefore, we examined ERK-MAPK activity within human epidermal keratinocytes in situ.
  
  
  RESULTS
  We used confocal microscopy and immunofluorescence labelling to measure the relative abundances of Raf-1, MEK1/2 and ERK1/2, and their phosphorylated (active) forms within three human skin samples. Additionally, we measured the abundance of selected proteins thought to modulate ERK-MAPK activity, including calmodulin, β1 integrin and stratifin (14-3-3σ); and of transcription factors known to act as effectors of ERK1/2, including the AP-1 components Jun-B, Fra2 and c-Fos. Imaging was performed with sufficient resolution to identify the plasma membrane, cytoplasm and nucleus as distinct domains within cells across the epidermis. The image field of view was also sufficiently large to capture the entire epidermis in cross-section, and thus the full range of keratinocyte differentiation in a single observation. Image processing methods were developed to quantify image data for mathematical and statistical analysis. Here, we provide raw image data and processed outputs.
  
  
  CONCLUSIONS
  These data indicate coordinated changes in ERK-MAPK signalling activity throughout the depth of the epidermis, with changes in relative phosphorylation-mediated signalling activity occurring along the gradient of cellular differentiation. We believe these data provide unique information about intracellular signalling as they are obtained from a homeostatic human tissue, and they might be useful for investigating intercellular heterogeneity.
authors.affiliation: Systems Biology Laboratory, Melbourne School of Engineering, University of Melbourne, Parkville, VIC Australia, 3010 ; ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of Melbourne, Parkville, Australia, 3010.
authors.forename: Joseph
authors.initials: J
authors.lastname: Cursons
authors.affiliation: Maurice Wilkins Centre, University of Auckland, Auckland, New Zealand ; School of Biological Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Angel
authors.affiliation: Systems Biology Laboratory, Melbourne School of Engineering, University of Melbourne, Parkville, VIC Australia, 3010.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Hurley
authors.affiliation: Maurice Wilkins Centre, University of Auckland, Auckland, New Zealand ; School of Biological Sciences, University of Auckland, Auckland, New Zealand ; Bioinformatics Institute, University of Auckland, Auckland, New Zealand ; Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: Cristin G
authors.initials: CG
authors.lastname: Print
authors.affiliation: Maurice Wilkins Centre, University of Auckland, Auckland, New Zealand ; School of Biological Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: P Rod
authors.initials: PR
authors.lastname: Dunbar
authors.affiliation: Department of Biology, New Zealand International College, ACG New Zealand, Auckland, New Zealand.
authors.forename: Marc D
authors.initials: MD
authors.lastname: Jacobs
authors.affiliation: Systems Biology Laboratory, Melbourne School of Engineering, University of Melbourne, Parkville, VIC Australia, 3010 ; ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of Melbourne, Parkville, Australia, 3010 ; School of Mathematics and Statistics, University of Melbourne, Parkville, Australia, 3010 ; School of Medicine, University of Melbourne, Parkville, Australia, 3010.
authors.forename: Edmund J
authors.initials: EJ
authors.lastname: Crampin
mesh_terms: D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005453:Fluorescence; D051921:Fos-Related Antigen-2 / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015603:Keratinocytes / Q000378:metabolism*; D048369:MAP Kinase Kinase 1 / Q000378:metabolism; D020935:MAP Kinase Signaling System; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D051921:Fos-Related Antigen-2; C581727:JunB protein, human; D016760:Proto-Oncogene Proteins c-fos; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D048049:Extracellular Signal-Regulated MAP Kinases; D048052:Mitogen-Activated Protein Kinase 3; D048369:MAP Kinase Kinase 1
keywords: Calmodulin; Cellular heterogeneity; Confocal microscopy; Homeostatic tissue; Immunofluorescence; Interfollicular keratinocytes; MEK1/2; Skin