Fig. 1: Perturbation of RNF40 influences mammary epithelial stem cell properties in vivo and in vitro.
From: RNF40 epigenetically modulates glycolysis to support the aggressiveness of basal-like breast cancer
A Venn diagram of human gene-regulatory E3 ligases (left panel) and their association with the stemness score in normal mammary epithelial (ITGA6+/EPCAM-) and BLBC (ITGB1+/ITGB41+) biopsies. B Schematic representation of the two transgenes of the MMTV-cre; Rnf40flox mouse model. C, D Hematoxylin and eosin staining and immunohistochemical detection of RNF40 and H2Bub1 on mammary gland sections from MMTV-cre; Rnf40fl/fl mice and MMTV-cre mice. Yellow arrows indicate mammary epithelial devoid of RNF40 and H2Bub1. Black scale bars: 50 μm. E Whole mounts staining of mammary glands showing a significant decrease of mammary duct branching density in MMTV-cre; Rnf40fl/fl mice compared to the control group with representative brightfield pictures (left panel) and the respective quantification (right panel). F, G Mammosphere and colony formation assay of isolated mammary epithelial cells from MMTV-cre; Rnf40fl/fl mice and their control group. H Organoid culture of murine mammary epithelial cells (right panel) showed a significant decrease of budding upon shRNF40 treatment compared to the control counterpart. White scale bar: 100 µm. I Colony formation assay of siControl- and siRNF40-treated MCF10A cells. E-I *p-val<0.05, **p-val<0.01, ***p-val<0.005. Statistical test: E, F, H (right panel), G–I: Student t-test; H (left panel): Mann Whitney test. Error bars: Standard error of the mean (SEM). All experiments were performed in at least three biological replicates per condition.
