Fig. 7
From: Single cell polarity in liquid phase facilitates tumour metastasis
MCAM regulates metastatic seeding in vivo. a Experimental metastasis assays of B16-F0 cells expressing no construct (ctrl.), GFP, Mcam or McamΔKKGK (ΔKKGK). The number of nodules on the lung was quantified after 2 weeks (all values normalised to mean of control, n = 8, mean ± SD, unpaired t-tests). Representative macroscopies (scale bar: 1 cm) and ki67-stained histological images of tumours (scale bar: 100 µm) are shown to the right. Mcam expression, polarity, transmigration and growth assays are shown in Supplementary Fig. 9a–d. b Metastatic seeding assays of B16-F0 cells expressing no construct (ctrl.), GFP, Mcam or McamΔKKGK (ΔKKGK). Seeding to the lung after 30 min was assessed by quantification of melan A+area per total area on histological stainings from mouse lungs. Representative gp100 stainings are shown to the right. Scale bar: 50 µm. (n = 6–10, mean ± SD, unpaired t-tests). c Experimental metastasis assays of B16-F1 cells expressing no shRNA (ctrl.), n.t. or Mcam-targeting (MCAM kd M1, MCAM kd M2) shRNAs quantified as in a (all values normalised to mean of control, n = 12–17, mean ± SD, unpaired t-tests). Representative macroscopies (scale bar: 1 cm) and ki67-stained histological images of tumours (scale bar: 100 µm) are shown to the right. Mcam expression, polarity, transmigration and growth assays are shown in Supplementary Fig. 9e–h. d Metastatic seeding assays of B16-F1 cells expressing no shRNA (ctrl.), n.t. or Mcam-targeting (MCAM kd M1, MCAM kd M2) shRNAs performed and quantified as described in b (n = 5–11, mean ± SD, unpaired t-tests). Representative gp100 stainings are shown to the right. Scale bar: 50 µm. (mean ± SD, unpaired t-tests). e Representative images of MCAM-stained primary melanoma samples showing absence (I) or MCAM expression in <50% of tumour cells (II), melanoma lymph node metastasis showing MCAM expression in >50% (III) and brain metastases (IV) showing MCAM expression in >90% of melanoma cells. Scale bars: 100 µm. f Evaluation of MCAM expression in samples of human primary and metastatic melanoma described in Supplementary Table 3 (Pearson’s Chi-squared test)
