Figure 2: TRB3 promotes tumour development in T2D mice.
(a) Blood glucose/insulin/IGF-1 was measured in C57 BL/6 and KK-Ay mice. Dashed lines indicate the normal range of blood glucose (3.5–9.8 mM) or insulin (0.2–0.6 ng ml−1). Data are mean±s.e.m. of three assays (n=8 per group). Inserts show TRB3 expression in the liver and lungs. (b) KK-Ay or C57 BL/6 mice were s.c. injected with B16-F10 cells (1.5 Ă— 105). Data are the mean volumes±s.e.m. at indicated times and representative mice (n=7 per group). Insert shows TRB3 expression in xenograft tumour. (c) Photographs of representative tumour and quantified tumour weight. Data are mean weight±s.e.m. (n=7 per group). Scale bar, 1 cm. (d) KK-Ay or C57 BL/6 mice were i.v. injected with B16-F10 cells (3 Ă— 105). Data are representative of macroscopy and bioluminescence images with total tumour volumes at multiple metastatic sites (mean±s.e.m.; n=12 per group). (e) Male KK-Ay (n=11) and C57 BL/6 (n=9) mice were p.o. gavaged with 0.1 ml (total 1 mg) DMBA once a week for 5 weeks. Data are timeline (upper) and macroscopic, and histopathological analysis of tumours (lower) with numbers of tumour nodules (mean±s.e.m.) and expression of TRB3 and γH2AX in the liver and lung. Scale bar, 10 μm. (f) Kaplan–Meier survival curve for mice fed with DMBA (n=20 per group). Statistical significance was determined by Kaplan–Meier log-rank test; *P<0.05. (g) Overexpression of TRB3 promotes DEN-induced tumorigenesis. Data are timeline and TRB3 expression in the liver and lungs (upper), and haematoxylin and eosin-stained sections plus incidence and number of tumour nodules (lower). Data are mean±s.e.m. (n=10 per group). Scale bar, 10 μm. (h) Blood glucose/insulin/IGF-1 was measured in mice infected with control-shRNA or TRB3-shRNA lentiviral particles (2 Ă— 105). Data are mean±s.e.m. of three assays with triplicates (n=8 per group). The inserts show TRB3 expression in the liver and lung. (i,j) TRB3 knockdown (n=9) or control KK-Ay (n=8) were s.c. inoculated with B16-F10 cells (1.5 Ă— 105). Data are the mean volumes±s.e.m. at indicated times and representative mice (i) and tumours along with tumour weight (j). Scale bars, 1 cm. (k) TRB3 knockdown (n=9) or control KK-Ay (n=8) were i.v. injected with B16-F10 cells (3 Ă— 105). Data are representatives of macroscopy and bioluminescence images with total tumour volumes (mean±s.e.m) at multiple metastatic sites. Except for f, statistical significance was determined with Student’s t-test; *P<0.05; **P<0.01; ***P<0.001.
