Fig. 7.

Notch1 signaling-dependent inhibitory effects of apoptotic lung cancer cell injection on CAF activation and metastasis in mice. a Schematic of the experimental design. Apoptotic 344SQ cells (ApoSQ) were subcutaneously injected into the established skin lesions 2 days after subcutaneous injection of 344SQ cells into syngeneic (129/Sν) mice (n = 9 per group). Where indicated, LY3039478 (8 mg/kg) or vehicle (Veh; 15% sugar gel) was orally administered three times a week for 6 weeks starting the day before injection of ApoSQ (n = 9 per group). Mice were necropsied 6 weeks after 344SQ cell injection. b Representative images of lungs with or without metastatic tumor nodules (yellow dashed circles). c Scatter plot of the numbers of lung metastatic nodules. NS: not significant, *P < 0.05, **P < 0.01, Kruskal‒Wallis test with Dunn’s post hoc test. The data are from representative images (b) or from independent experiments with 9 mice per group (mean ± standard error; c). d Numbers of mice with (w/) and without (w/o) visibly determined metastases (Met). e qRT‒PCR analysis of CAF markers, MMPs, growth factors (GFs)/chemokines, and Notch1-related molecules in Thy1⁺ CAFs isolated from primary tumors. NS: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal‒Wallis test with Dunn’s post hoc test. The data are from three replicates per condition, with cells pooled from three mice per replicate. The data are shown as the means ± standard errors