Fig. 2

COMMD3 low expression correlates with poor overall survival in breast cancer patients. A. T-47D cells were transduced with pGIPz shControl or representative shCOMMD3 hairpin (V2LHS_28871) and selected for 48 h. Cells were plated on GFR matrigel in 2% FCS and grown for 14 days prior to imaging. B. Box plot depicting acini size upon COMMD3 depletion measured on day 14; n = 2, more than 100 acini were counted. C. Analysis of COMMD3 mRNA levels in FACS-sorted subtypes of human mammary epithelium, taken from Lim et al. (36). Basal/MaSC – basal and stem cells; Lumpro – luminal progenitor cells; Lum – mature luminal cells; Stroma – stromal cells (including fibroblasts). D. Box plot depicting COMMD3 mRNA expression in TCGA breast cancer samples, grouped according to PAM50 subtype (http://tumoursurvival.org). N-Like is normal-like. E. Kaplan–Meier survival analysis of the relationship between COMMD3 mRNA expression and breast cancer patients' clinical outcome using the TCGA RNAseq dataset. COMMD3 expression stratified overall survival. F. Representative images of COMMD3 antibody optimisation by immunohistochemistry (IHC) in HEK293T cells after transfection with the shRNA COMMD3 or control vector. G. Representative images of COMMD3 expression in breast cancer tissues on the TMA. Examples of 0–4 + TMA core staining are shown. H. Kaplan–Meier survival analysis of the relationship between COMMD3 protein levels and outcome for ER + HER2-negative tumours (luminal A with low Ki67 staining) on the TMA. COMMD3 expression stratified overall survival. I. Quantification of COMMD3 protein association with tubule formation, ELF5, c-Kit and AR staining that were assessed for this human breast cancer TMA previously [25,26,27,28,29].