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

固定单细胞的细胞内(磷酸化)蛋白质和转录组学的联合定量

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作者:Jan P Gerlach, Jessie A G van Buggenum, Sabine E J Tanis, Mark Hogeweg, Branco M H Heuts, Mauro J Muraro, Lisa Elze, Francesca Rivello, Agata Rakszewska, Alexander van Oudenaarden, Wilhelm T S Huck, Hendrik G Stunnenberg, Klaas W Mulder

Abstract

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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