Fig. 1
From: Proliferative heterogeneity of murine epithelial cells in the adult mammary gland
Single cell gene expression analysis of cycling mammary epithelial cells. a Immunofluorescence staining for mCherry to detect cells in G1 (red; arrows) and mVenus to detect S/G2/M (green; arrowheads) phases of the cell cycle in intact mammary glands. ERα (cyan) marker was used to discriminate between the HR− (yellow arrow/arrowheads) and HR+ (white arrow/arrowheads) luminal cells. Scale bar = 25 µm. b Schematic of gating strategy for single cell mammary epithelial cell analysis. c Heat maps showing unsupervised hierarchical clustering of gene expression of cells in G1, G1hi, S/G2/M and DN of the cell cycle, of 96 single cells from each of the basal, HR− and HR+ luminal compartments. Coloured symbols above heat map correspond to the cell cycle population. White to red intensity suggests middle to high expression, whereas white to blue suggests low to no expression. Violin plots show the distribution of cell cycle/mammary-related gene expression of single cells from the different cell cycle phases of the (d) basal, (e) HR− luminal and f HR+ luminal populations. Two-way ANOVA analysis with Tukey’s multiple comparison test was used to determine gene expression changes between different cell cycle phases. *, **, *** represent statistical significant at P < 0.05, P < 0.01, P < 0.001, respectively. No asterisks, statistically not significant
