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. 2016 Aug 5;7:12321. doi: 10.1038/ncomms12321

Figure 2. Reduced Hh signalling increases tumour burden in the AOM/DSS model.

Figure 2

(a) Schematic of Tam and AOM/DSS treatment of IhhΔVil mice. (b) Tumour numbers and tumour burden for IhhΔVil mice (n=13) and controls (n=11 littermates that lacked either the VillinCreER allele or floxed Ihh alleles or both): 5/13 IhhΔVil mice and 11/11 controls reached the endpoint; P=0.0321 (tumour numbers), P=0.0215 (tumour burden; both t-tests). (c) Representative confocal image of ZsGreen reporter fluorescence in an IhhΔVil mouse (upper left panel). Persistent reporter expression indicates enduring loss of Ihh in the tumour. Haematoxylin and eosin (H&E) staining of consecutive section (upper right panel). Littermate wt controls are shown in the lower panels. Dashed boxes show magnified areas. Nuclear counterstain: TOPRO-3. Scale bars, 1 mm/250 μm. Representative image of n=2 IhhΔVil mice with n=8 tumours, in which all tumours expressed ZsGreen, whereas no zsGreen expression was detected in n=3 control mice. (d) Real-time quantitative PCR (RT-qPCR) for the indicated transcripts in IhhΔVil mice (n=5) and controls (n=5); P=0.003 (Ihh), P=0.887 (Shh), P=0.311 (Gli1), P=0.0142 (Ptch1; all t-tests, P-values after multiple test correction (fdr)). Bars represent mean and whiskers represent s.e.m. (e) Schematic of tumour induction, vismodegib treatment and tumour quantification with US; wt C57BL/6 mice received either 25 mg kg−1 vismodegib or vehicle (DMSO) twice daily, 6 days per week. (f) Tumour numbers and tumour burden for mice treated with vehicle (n=16) or vismodegib (n=15): 15/16 DMSO-treated and 10/15 vismodegib-treated mice reached the endpoint; P=0.0014 (tumour numbers), P=0.0036 (tumour burden; both t-tests). (g) Representative histology of tumours from the indicated treatment groups (H&E); scale bars, 1 mm/200 μm. *P<0.05, **P<0.01 and ***P<0.001.