Figure 1: Single-cell RT-PCR analysis in freshly isolated vascular cells. | Nature Communications

Figure 1: Single-cell RT-PCR analysis in freshly isolated vascular cells.

From: Single-cell profiling reveals heterogeneity and functional patterning of GPCR expression in the vascular system

Figure 1

EC or SMC obtained from skeletal muscle vasculature (sk), lung (lu), aorta (ao) of Cdh5-Crepos; dTom/EGFP-reporterpos mice (Cdh5-EGFP) and tamoxifen-treated Myh11-CreERT2pos; dTom/EGFP-reporterpos mice (Myh11-EGFP), respectively, were subjected to single-cell RT-PCR. (a) Expression of identity-defining genes and quality control genes (Gapdh, Hprt and Actb) after exclusion of contaminating cells or marker negative cells (each dot one cell). (b) Comparison of expression data obtained by multiplex RNA expression analysis in pooled SMsk and single-cell RT-PCR (sc RT-PCR) in individual SMsk. GPCRs are arranged on the abscissa according to their expression strength in pooled RNA analysis, the ordinate shows the strength of gene expression in individual cells (each dot one cell). Green boxes indicate genes negative in sc RT-PCR but positive in pooled RNA analysis (cell/animal numbers: sc RT-PCR: n=57 cells from seven mice; pooled RNA analysis: 500 ng from 106 cells per six mice. Values of genes that are not expressed were scattered around 0 to allow graphical estimation of cell counts). Expression data are calculated as follows: Gene expression=2(Limit of detection(LoD) Ct—sample Ct); LoD Ct is set to 24.

Back to article page