Figure 1. Generation of Wnt/iFGFR^GFP tumors to study cell signaling and tumor microenvironment during dormant MRD and recurrence.

A) Workflow of tumor generation for various stages including primary, dormant, and recurrent. Activation of iFGFR1 and growth of transplanted Wnt/iFGFR^GFP tumors were stimulated by dimerizer treatment. Once primary tumors reached 8–10mm in diameter, mice were treated with BGJ398 for 14 days. Regressed tumors (image Ai and Aii showing GFP expression) were off drug treatment for another 14 days and then collected. A separate cohort was used to study recurrence after 1–4 months from BGJ398 treatment. Tumors were collected when they reached 1.5cm in diameter both at the primary and recurrent stage. B) Growth curve for “recurrent” tumors. Tumors were designated as “recurrent” when they reached 5mm in diameter. Three lesions that remained dormant for more than 6 months were harvested and labeled long-term DT. C) Survival graph showing mice with recurrent tumors same as in B (12 out of 15 total mice). The 3 long-term DT lesions were collected without recurrence. D) H&E staining, D) Immuno-fluorescent (IF) staining of Ki67 and GFP, and E) Immuno-histochemistry (IHC) staining of phosphorylated-p27 (p-p27) at different stages.