Figure 6: Effect of TGF-β signalling on dormancy and self-renewal markers.
(a) Immunblots for the indicated antigens on PT-HEp3 (PT), BM-D1 and BM-T1 cell lysates after TGF-β2 (2 ng ml−1) or TGF-β-RI inhibitor (LY-364947, 5 μM) treatment for 24 h in SF media. (b) Immunoblots for the indicated antigens on T-HEp3 cell lysates after treatment with TGF-β2 (2 ng ml−1) for 24 h in SF media. (c,d) qPCR-measured DEC2 (c) and p53 (d) mRNA levels in the indicated cells after treatment with 2 ng ml−1TGF-β2 or LY-364947, 5 μM for 24 h in SF media (n = 6 RNA samples were assessed over 3 independent experiments). (e) BM-D1 growth on CAMs for 4 days after TGF β 2 knockdown. BM-D1 cells were transfected with either scrambled (scr) or TGF β 2 siRNA and inoculated into CAMs (5×105 cells per CAM) for 4 days (n = 20 (scr), 19 (TGF β 2) tumours per condition). (f) BM-D1 growth on CAMs for 4 days after TGF-β-RI inhibition with LY-364947 (5 μM). BM-D1 cells were pre-treated with TGF-β-RI inhibitor (LY-364947, 5 μM) for 24 h in SF media and then inoculated into CAMs (5×105 cells per CAM) for 5 days. Tumour nodules were treated with LY-364947 (5 μM) every day in the CAMs (n = 9 tumours per condition). Data in c,d represent mean±s.e.m. In c–f, *P<0.05, ***P<0.001 by Mann–Whitney test. Uncropped images of blots are shown in Supplementary Fig. 6.
