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

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 diffe...

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Autores principales: Gerlach, Jan. P., van Buggenum, Jessie A. G., Tanis, Sabine E. J., Hogeweg, Mark, Heuts, Branco M. H., Muraro, Mauro J., Elze, Lisa, Rivello, Francesca, Rakszewska, Agata, van Oudenaarden, Alexander, Huck, Wilhelm T. S., Stunnenberg, Hendrik G., Mulder, Klaas W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6365588/
https://www.ncbi.nlm.nih.gov/pubmed/30728416
http://dx.doi.org/10.1038/s41598-018-37977-7
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author Gerlach, Jan. P.
van Buggenum, Jessie A. G.
Tanis, Sabine E. J.
Hogeweg, Mark
Heuts, Branco M. H.
Muraro, Mauro J.
Elze, Lisa
Rivello, Francesca
Rakszewska, Agata
van Oudenaarden, Alexander
Huck, Wilhelm T. S.
Stunnenberg, Hendrik G.
Mulder, Klaas W.
author_facet Gerlach, Jan. P.
van Buggenum, Jessie A. G.
Tanis, Sabine E. J.
Hogeweg, Mark
Heuts, Branco M. H.
Muraro, Mauro J.
Elze, Lisa
Rivello, Francesca
Rakszewska, Agata
van Oudenaarden, Alexander
Huck, Wilhelm T. S.
Stunnenberg, Hendrik G.
Mulder, Klaas W.
author_sort Gerlach, Jan. P.
collection PubMed
description 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.
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spelling pubmed-63655882019-02-08 Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells Gerlach, Jan. P. van Buggenum, Jessie A. G. Tanis, Sabine E. J. Hogeweg, Mark Heuts, Branco M. H. Muraro, Mauro J. Elze, Lisa Rivello, Francesca Rakszewska, Agata van Oudenaarden, Alexander Huck, Wilhelm T. S. Stunnenberg, Hendrik G. Mulder, Klaas W. Sci Rep Article 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. Nature Publishing Group UK 2019-02-06 /pmc/articles/PMC6365588/ /pubmed/30728416 http://dx.doi.org/10.1038/s41598-018-37977-7 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Gerlach, Jan. P.
van Buggenum, Jessie A. G.
Tanis, Sabine E. J.
Hogeweg, Mark
Heuts, Branco M. H.
Muraro, Mauro J.
Elze, Lisa
Rivello, Francesca
Rakszewska, Agata
van Oudenaarden, Alexander
Huck, Wilhelm T. S.
Stunnenberg, Hendrik G.
Mulder, Klaas W.
Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
title Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
title_full Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
title_fullStr Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
title_full_unstemmed Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
title_short Combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
title_sort combined quantification of intracellular (phospho-)proteins and transcriptomics from fixed single cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6365588/
https://www.ncbi.nlm.nih.gov/pubmed/30728416
http://dx.doi.org/10.1038/s41598-018-37977-7
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