Fig. 5

UNC5A knockdown results in luminal/basal hybrid and bipotent luminal progenitor phenotype. a ΔNp63 levels are significantly elevated in sh-UNC5A MCF7 cells. Cells were treated with vehicle or estradiol (E2) for 3 h and qRT-PCR was used to measure ΔNp63 and TAp63 levels (mean ± SEM, n = 2). Data were analyzed as in Fig. 3e (*p < 0.05, **p < 0.01, and ***p < 0.001). TP63 protein levels in sh-Control and sh-UNC5A MCF7 cells treated with or without E2 for 24 h are shown on the right. TP63 is expressed as multiple isoforms and there appears to be isoform switching in sh-UNC5A cells compared with sh-Control cells. b ΔNp63 and TAp63 levels in sh-Control and sh-UNC5A T-47D cells. c sh-UNC5A cells have lower ELF5 mRNA compared with sh-Control cells. Cells were treated with vehicle or E2 for 3 h and qRT-PCR was used to measure ELF5. d UNC5A knockdown leads to elevated NTN4. e UNC5A knockdown leads to elevated MECOM (EVI-1) expression in T-47D cells. Although MECOM levels were elevated in MCF7 cells upon UNC5A knockdown (Additional file 6), proteins were not detected by Western blotting. f UNC5A knockdown T-47D cells express both KRT14 (basal; green) and KRT19 (luminal; red) while sh-Control T-47D cells express luminal KRT19. Results for TMCF7 cells are shown in Additional file 8. g sh-UNC5A MCF7 cells form irregularly shaped mammospheres. h UNC5A knockdown MCF7 cells display phenotypic characteristics similar to luminal progenitor (CD49f⁺/EpCAM⁺)/cancer stem cells (CD44⁺/CD24⁻) compared with sh-Control cells. A histogram displaying CD49f and CD44 expression in different cell types is depicted on the right