Tumor control is largely preserved despite FTY720 administration at later time points. B16.SIY-bearing mice were either treated with FTY720 or control vehicle prior to tumor inoculation (B), prior to initiation of therapy (C) or with the last dose of αCTLA-4 mAb on day 10 (D). A. Schematic overview of the experimental design bold arrow heads indicate initiation of FTY720 treatment, which was always carried out through the entire experiment. B-D. Tumor outgrowth measured in mm2 comparing vehicle control (circle) to FTY720 treatment (square) on no treatment (open) vs. αCTLA-4 + αPD-L1 (filled). B. FTY720 treatment was initiated 24 h prior to tumor inoculation. C. FTY720 treatment was given on day 4, 2.5 h before therapy was initiated. D. FTY720 was given at the same time point as the last dose of αCTLA-4 mAb treatment. Depicted is a representative experiment, showing mean of 5 mice +/− SEM. Significance was determined using a two-way ANOVA with Bonferroni post-test.