Figure 3

DUOX1 inhibition autonomously reduces aggressiveness and growth of transplanted SKCMs. (A) Schematic diagram showing adult allotransplantation procedure. One-year-old Casper zebrafish were irradiated 2 days before transplantation to prevent tumor rejection. Three hundred thousand cells from the nodular tumors of either MinicoopR mitfa:NRAS-QW61R/mitfa:DN-DUOX1 or MinicoopR mitfa:NRAS-Q61R/mitfa:EGFP (control) fish were subcutaneously injected in pre-irradiated recipients, and images were taken weekly during the following 4 weeks after transplantation and analyzed as indicated in the Methods and Materials section. Arrow, timeline; dpi, days post-injection; wpt, weeks post-transplant. (B) Representative images of transplanted melanoma growth rate of MinicoopR mitfa:NRAS-QW61R/mitfa:DN-DUOX1 and MinicoopR mitfa:NRAS-Q61R/mitfa:EGFP in pre-irradiated adult Casper zebrafish. (C) Average tumor size for each week post-transplant. Each dot represents a recipient-transplanted fish, and the mean is also shown. ** p < 0.01, **** p < 0.0001 according to unpaired Student t test. (D) Growth rate of transplanted MinicoopR mitfa:NRAS-QW61R/mitfa:DN-DUOX1 and MinicoopR mitfa:NRAS-Q61R/mitfa:EGFP SKCMs. DN-DUOX1: n = 5 tumors and 111 recipient fish; EGFP: n = 3 tumors and 72 recipient fish.