Fig. 6
Expression of neural markers, Gpr158 and Ccnd1 in allografts of miR-449ahigh and miR-449alow tumours. Phenotype of tumours generated from allografted Rb/p53 (left column), miR-449a antagomir-treated Rb/p53ant (centre column) and Pten/p53 cells (right column) into NOD-SCID mice. a Morphology (H&E) shows a well demarcated tumour border in Rb/p53 PNET (single line of arrowheads), and an infiltrative margin in both, antagomir-treated Rb/p53 grafts (b) and Pten/p53 grafts (c). Image analysis of regions of interest on whole slide digitized images of immunostained histological sections with Definiens Developer shows a reversal of Rb/p53ant grafts to levels found in Pten/p53 grafts (markers Olig2 (d–g), Pdgfrα (l–o), and Sox2 (p-s), whilst only DCX it is unaltered (h–K). Expression of these markers is reduced in Rb/p53; miR-449ahigh tumours, high in Pten/p53 (miR-449alow) grafts, and restored upon suppression of miR-449a in antagomir-treated Rb/p53ant; miR-449alow grafts. (t-ae) miR-449a regulates Ccnd1 and GPR158 in tumours in vivo: (t–w), Ccnd1 expression is low in Rb/p53 (miR-449ahigh) tumours (t), and high in Pten/p53 (miR-449alow) tumours (u) as well in antagomir-treated Rb/p53; miR-449alow tumours (v, w). Gpr158 expression is regulated correspondingly (x–aa). In keeping with the shorter survival of mice with Rb/p53; miR-449ahigh tumours, proliferation is high in these tumours (ab). Scale bar 50 µm. All figures: *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001 (Student’s t-test). Each value represents the mean ± s.d
