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Figure 1 | Breast Cancer Research

Figure 1

From: Mouse mammary stem cells express prognostic markers for triple-negative breast cancer

Figure 1

Isolation of high-purity basal mammary stem cells. (A, B). Flow cytometry of mouse mammary epithelial cells isolated from 8- to 10-week-old virgin FVB mice and stained with antibodies against CD24, Sca-1, CD49f and c-Kit. (A) Isolation of the luminal ER- progenitor (Gate I) and luminal ER+ cell populations (Gate II) on the basis of CD24 and Sca-1 expression followed by c-Kit as previously described [13]. (B) Isolation of basal myoepithelial (MYO; Gate III) and mammary stem cells (MaSCs; Gate IV), from the same sort data as in (A), on the basis of CD24 and CD49f expression followed by c-Kit as previously described [13]. Only epithelial cell data is shown. For the full gating hierarchy see Additional file 1. (C) qrtPCR gene expression analysis of mammary epithelial lineage-specific genes. Data expressed as mean fold expression (±95% confidence intervals) over comparator population (luminal ER- cells) in three independent isolates of each cell population. Statistical significance was determined according to [74]. ** P <0.01 relative to comparator. ‘Typical expression’ indicates the populations expected to have the highest levels of expression of the genes based on previous findings [13]. § Dmbt1 expression was undetectable in all isolates of MaSC and MYO populations. (D) Results of low-dose transplantation of mammary subpopulations into cleared fat pads. The number of successful outgrowths, the total number of fat pads transplanted and the extent each outgrowth filled the fat pads, as indicated by the pie chart symbols, are shown. ER, estrogen receptor; qrtPCR, quantitative real-time rtPCR.

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