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. 2018 Dec 13;12(1):135–151. doi: 10.1016/j.stemcr.2018.11.010

Figure 2.

Figure 2

Dach1 RNA Transcripts in Mammary Epithelial Cells during Development

(A) Diffusion map of single-cell RNA sequencing (scRNA-seq) transcriptomes annotated by stage of development. Epcam+ mammary epithelial cells (MECs) were derived from developing (embryonic day 16 [E16] and E18), post-natal day (P4) and adult MECs (Giraddi et al., 2018), and scRNA-seq was plotted according to diffusion components (DCs) using software (http://uofuhealth.utah.edu/huntsman/labs/spike/d3.php) as described previously (Giraddi et al., 2018). DC1 represents a continuum of relative enrichment from basal to luminal subtype. DC2 represents developmental time from primitive (E16 and E18) to P4 adult mammary cells occupy ends of the DC2 axis.

(B) The abundance of Dach1 transcripts was mapped upon the diffusion map scRNA-seq transcriptomics, in order to show the correlation with stage of development and relative abundance in the luminal versus basal subtypes. Dach1 is expressed among multipotent fetal mammary stem cells in the embryo and among mixed lineage precursors prior to puberty Dach1 expression was detected in adult basal cells (dark blue dots in lower-right corner) and (ERα+) luminal progenitors (light blue dots in upper-right corner).

(C–F) Dach1 RNA transcripts are expressed during post-natal development. (C) scRNA-seq of MECs derived from nulliparous, mid gestation, lactation, and post-involution mammary glands identified 15 clusters of MECs (Bach et al., 2017). (D) Relative expression of Dach1 within each cluster type is shown colorimetrically, with relative expression shown in the bar to the right. (E) Dendrogram of clusters based on the log-transformed mean expression values of the 15 clusters. The red blocks show the cluster subtype in which Dach1 is expressed based on the relative abundance of transcripts shown in (F). Dach1 gene expression was detectable primarily in presumptive basal and myoepithelial cells (clusters C12, C13, and C14), and to a lesser degree also in luminal ERα+ (cluster C3) and luminal progenitor clusters (C6/7).