a, IL12-alb-lum produced by LNP-replicon-transfected tumor cells binds to collagen I. B16F10 cells were transfected with LNP-Rep(IL-12-alb-lum) and the supernatants of the transected cells were added to collagen-coated plates for analysis of binding by ELISA in comparison of standards protein IL12-alb-lum. Shown are the ELISA absorbance for IL-12 detection versus concentration of added IL-12-alb-lum using the supernatants of B16F10 cells that were transfected with LNP-Rep(IL-12-alb-lum) or left untreated (n = 4 technical cell culture replicates from one of two independent experiments. b, c, Comparison of AST (b) and ALT (c) levels in serum at day 1 and 3 after the indicated treatments (error bars are mean + s.e.m. from n = 5 mice/group). d, Comparison of CXCL9 and CXCL10 levels in tumors at day 1 and 3 after the indicated treatments (error bars are mean + s.e.m. from n = 5 mice/group). P values were determined by two way ANOVA analysis using PRISM Software and exact P values were indicated.