Fig. 3
Combination therapy with doxorubicin leads to therapeutic immunity and systemic effects. A Schematic representation of the experimental setup. Treatment was started when large established tumors were reached (120–150 mm3). B, C Figures show 4T1 tumor growth of individual mice in each group (B) as well as survival (C) (one experiment was performed with n = 12/group). D Schematic representation of the experimental setup for a two-site tumor model. Primary tumors were inoculated at the right flank followed by a second tumor inoculation at the contralateral flank. Primary tumors were p.l. treated whereupon systemic effects of local treatment were analyzed on disseminated non-treated tumors (left flank). E The graphs show s.c. 4T1 tumor volume of individual values ± SEM at day 18 (n = 5). F Tumor growth curves of both treated (upper panel) and non-treated (lower panel) tumors of individual mice using the s.c. 4T1 mammary carcinoma model. Mice were treated with PBS (grey), doxo (black), 30 μg of Bisp-AFN Clec9A-PDL1-AFN (green) or a combination of Clec9A-PDL1-AFN with doxo (blue). Black lines underneath the X-axis depict the treatment time, while orange arrows indicate s.c. administration of doxo. Kaplan Meier plots depicting % survival (C) were analyzed using log-rank (Mantel-Cox) test. Bar plots (E) were analysed using One-way ANOVA Kruskal–Wallis test with Dunn’s multiple comparisons test (treated tumors) or using One-way ANOVA followed by bonferroni’s multiple comparison test (non-treated tumors) dependent on Shapiro–Wilk test for normal distribution of the data. * < 0.05; ** < 0.01; *** < 0.001; **** < 0.0001