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. 2015 Jul 31;16(9):1145–1163. doi: 10.15252/embr.201540759

Figure 2.

Figure 2

Loss of BNip3 promotes early progression to carcinoma and increased lung metastases

  • A, B Primary mammary tumors in BNip3 null mice at d35 showed increased nuclear grade (grade 3) compared to wild-type (grade 2).
  • C, D Expression of estrogen receptor-alpha (ER-α) at d65 shows reduced expression in BNip3 null tumors compared to wild-type.
  • E, F Increased pleomorphism and evidence of epithelial–mesenchymal transition (EMT) in BNip3 null tumors at d80 but not in wild-type.
  • G, H Loss of basement membrane integrity in BNip3 null tumors at d65 (indicated by the red arrow).
  • I, J Laminin-α1 staining confirms loss of basement membrane integrity in BNip3 null tumors at earlier stages than for wild-type (black arrows indicate laminin-α1-expressing basement membrane in wild-type and loss of basement membrane integrity in BNip3 null).
  • K, L Migration–invasion assays in Transwell assays, measured in triplicate. Quantification of invasion through collagen-coated Transwell inserts and migration across uncoated inserts for wild-type and BNip3 null MECs grown under 20% or 1% oxygen.
  • M, N Growth of wild-type and BNip3 null 3D spheres in Matrigel, measured in triplicate.
  • O, P Increased lung metastases at d80 in BNip3 null mice (n = 21) compared to wild-type (n = 24).

Data information: Black scale bar is 50 μm. White scale bar is 20 μm. Results are expressed as the mean ± SEM. ***P < 0.001, ****P < 0.0001.