Fig. 2

TRPS1 controls a luminal progenitor transcriptional program a H & E stainings of mammary gland cross-sections from shRen and shTrps1 mice treated with Doxycycline for 10 weeks. Left: overview. Right: close-up of a mammary duct. scale bars = 50 µm. b Immunofluorescent stainings of the indicated proteins in mammary ducts from shRen and shTrps1 female mice treated for 10 days with doxycycline. Scale bar = 50 µm. c Representative flow cytometry plots of Luminal and basal cells expressing shRNAs (GFP⁺) d Principal Component Analysis (PCA) plots RNA-Seq data from Luminal and basal cells expressing shRen or shTrps1 e Volcano plots showing differential gene expression in luminal (left) and basal (right) cells expressing shTrps1 vs shRen. f Gene set enrichment analysis (GSEA) of shTrps1 vs shRen luminal cells using C6 oncogenic gene sets. g GSEA Enrichment plot for YAP signature [20] in shTrps1 vs shRen luminal cells. h Gene set enrichment analysis of shTrps1 vs shRen luminal cells using C2 curated gene sets. i GSEA Enrichment plot for Luminal progenitor signature [10] signature in shTrps1 vs shRen luminal cells j Overview of the genetic structure of the luminal-specific TRPS1 depletion mouse line. The line harbors two transgenes. One is composed of the Krt8 promoter and the CreERT2 recombinase coding sequence. The other contains rtTA3 and mKate2 coding sequences under the control of a CAG promoter. A Lox-STOP-Lox prevents their expression. In addition, a TRE-tGFP-shRNA element is inserted in the Col1a1 locus. Tamoxifen allows excision of STOP by recombinase and expression of rtTA3 and mKate2. Expression of tGFP and shRNA is enabled by rtTA3 in the presence of doxycycline. k Immunofluorescent stainings of the indicated proteins in mammary ducts from luminal-specific TRPS1-depletion mouse. Scale bar = µm. l) Representative flow cytometry plots of recombined (mKate2⁺) Luminal and basal cells expressing shRNAs (GFP⁺) m Volcano plots showing differential gene expression in luminal cells expressing shTrps1 vs shRen n GSEA Enrichment plot for Luminal progenitor signature [10] in shTrps1 vs shRen luminal cells