Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells.


Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
06 02 2019
Historique:
received: 20 09 2018
accepted: 14 12 2018
entrez: 8 2 2019
pubmed: 8 2 2019
medline: 19 8 2020
Statut: epublish

Résumé

Environmental stimuli often lead to heterogeneous cellular responses and transcriptional output. We developed single-cell RNA and Immunodetection (RAID) to allow combined analysis of the transcriptome and intracellular (phospho-)proteins from fixed single cells. RAID successfully recapitulated differentiation-state changes at the protein and mRNA level in human keratinocytes. Furthermore, we show that differentiated keratinocytes that retain high phosphorylated FAK levels, a feature associated with stem cells, also express a selection of stem cell associated transcripts. Our data demonstrates that RAID allows investigation of heterogeneous cellular responses to environmental signals at the mRNA and phospho-proteome level.

Identifiants

pubmed: 30728416
doi: 10.1038/s41598-018-37977-7
pii: 10.1038/s41598-018-37977-7
pmc: PMC6365588
doi:

Substances chimiques

Quinazolines 0
Tyrphostins 0
RTKI cpd 170449-18-0
Focal Adhesion Kinase 1 EC 2.7.10.2
PTK2 protein, human EC 2.7.10.2

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1469

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Auteurs

Jan P Gerlach (JP)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.
Department of Molecular Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Jessie A G van Buggenum (JAG)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Sabine E J Tanis (SEJ)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Mark Hogeweg (M)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Branco M H Heuts (BMH)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Mauro J Muraro (MJ)

Oncode Institute, Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands.

Lisa Elze (L)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Francesca Rivello (F)

Radboud University, Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ, Nijmegen, The Netherlands.

Agata Rakszewska (A)

Radboud University, Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ, Nijmegen, The Netherlands.

Alexander van Oudenaarden (A)

Oncode Institute, Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands.

Wilhelm T S Huck (WTS)

Radboud University, Institute for Molecules and Materials, Heyendaalseweg 135, 6525 AJ, Nijmegen, The Netherlands.

Hendrik G Stunnenberg (HG)

Department of Molecular Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.

Klaas W Mulder (KW)

Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Science Faculty, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands. k.mulder@science.ru.nl.

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Classifications MeSH