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. 2013 Jan 11;5(2):264–279. doi: 10.1002/emmm.201201320

Figure 2. The self-renewal marker HMGA2 is highly expressed in Wnt10bTG mammary tumours and mammary placodes.

Figure 2

  • A. IHC for oestrogen receptor α (ERα), progesterone receptor (PR) and Her2 to validate the triple-negative phenotype of Wnt10bLacZ tumours. Wild-type (WT) uterus and Her2 tumour samples were used as positive controls. Highlighted are myoepithelial (myo), glandular epithelial (ge), luminal epithelial (le) and stromal cells (s).
  • B. Hierarchical clustering of microarray data of Hmga2 and Hmga1 expression in Wnt10b-driven tumours.
  • C. Validation of Hmga2 and Hmga1 expression in tumour cells (Tu), Lin−LacZ+ (LacZ+) tumour cells compared to wild-type virgin (Virg) and ErbB2TG tumour cells, as determined by qt-PCR. Error bars represent the means and the standard deviations from three independent experiments; **p-value = 0.007 versus Virg or ErbB2TG Tu samples (Student's t-test).
  • D. IHC of HMGA2 in WT mammary glands, and ErbB2TG and Wnt10bLacZ tumours. Red arrowheads highlight cells positive for HMGA2 and black arrowheads negative cells, respectively.
  • E. IHC of HMGA2 in WT and Wnt10b knockout (Wnt10b−/−) mammary placodes (mp) at embryonic Day 14.5 reveals loss of HMGA2 expression in Wnt10b−/− mp. Dashed black lines indicate Wnt10b−/− mp. Mesenchymal (M: red arrow head), epidermis layer (EL: short black arrow) and mammary anlagen (MA: long black arrow).
  • F, G. Mammosphere assays (MSA) of WT and Wnt10b−/− mammary gland cells and MMTV-Wnt10bLacZ primary tumour cells that were serially passaged (at Day 10 and 20) and analysed at Day 30. Secondary spheres were counted in sequential dilutions and imaged using a phase contrast microscope. Biological replicates (n = 3) and six technical replicates are shown. Wnt10bLacZ-derived MSAs analysed by IHC for HMGA2 is shown (10×) in lower right panel of G (bar, 50 µm). In F error bars represent the means and the standard deviations from three independent experiments; p values: a = 0.04, b = 0.03 versus wild-type or Wnt10b−/− samples (Student's t-test). p Values of <0.05 were considered to be statistically significant (C, F).