Figure 2From: A pooled shRNA screen for regulators of primary mammary stem and progenitor cells identifies roles for Asap1 and Prox1Selection of candidate genes for further analysis. (A) Venn diagram summarizing expression of genes, as determined by RNA-Seq, targeted by shRNAs with FDR < 0.01 in freshly isolated MaSC/basal, LP and ML populations and MaSC/basal-derived mammosphere (Mammosphere) populations. (B) qRT-PCR profiling of two candidate regulators, Prox1 and Asap1, in primary epithelial subsets (n = 3; mean ± S.E.M). (C) Immunohistochemistry showing PROX1 and ASAP1 protein expression in mammary epithelial cells of 8-week-old virgin mice. (D) Representative FCM plots showing the relative abundance of MaSC/basal cells transduced with shControl-GFP, shProx1-GFP or shAsap1-GFP retrovirus and competitor cells transduced with control mCherry-expressing retrovirus at Day 2 and Day 7 in i3T3 cultures. (E) Histogram showing the change in the ratio of shRNA-GFP+: mCherry+ cells for each shRNA between 2 and 7 days of co-culture. Data from 3 independent experiments displayed as mean ± S.E.M. Statistical significance was calculated relative to shControl: shProx1, p ≤ 0.035; shAsap1, p ≤ 0.013.Back to article page