Fig. 1. Gene expression analysis of EPFs and ENFs.
(A) Schematic showing mTmG system. (B) Hierarchical clustering of simultaneous gene expression for at least 70 individual cells FACS-isolated as either unfractionated dermal lysate (blue), EPFs (green), or ENFs (red) in P56 En1Cre;R26mTmG mice. Gene expression is presented as relative (fold) change from median on a color scale from yellow (high expression, 32-fold above median) to blue (low expression, 32-fold below median). K-means clustering of cells demonstrates two “fibroblast” clusters and a smaller nonfibroblast population. (C) Differentially expressed genes between unfractionated dermal lysate cells, EPFs, and ENFs identified using nonparametric two-sample Kolmogorov-Smirnov testing (P < 0.01 after Bonferroni correction for multiple comparisons). Distributions of single-cell gene expression between populations are illustrated here using median-centered Gaussian curve fits. The left bar for each panel represents the fraction of qPCR reactions that failed to amplify in each group. (D) qRT-PCR analysis of fibroblast- and nonfibroblast-associated gene expression in unfractionated dermal lysate and FACS-isolated EPFs and ENFs.