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. 2018 Jun 28;9:1496. doi: 10.3389/fimmu.2018.01496

Figure 4.

Figure 4

Invariant natural killer T response in mice injected intratumorally with fd-vectorized alpha-GalactosylCeramide (α-GalCer). Mice were inoculated with B16 melanoma cells and when tumor was palpable, mice were intratumorally injected with free α-GalCer, α-GalCer delivered by phage particles or vehicle alone, and sacrificed after 24 h. (A) Schematic representation of the experiment schedule. (B) CD3+NK1.1+ cells were evaluated in freshly isolated spleen of injected mice by FACS analysis and staining with anti-NK1.1 and anti-CD3 antibodies. Percentage of NK1.1+CD3+ positive cells ± SD is reported. p < 0.01 by one-way analysis of variance (ANOVA) followed by a Dunnett’s multiple comparison test. **p < 0.01. (C,D) Splenocytes were cultured with graded doses of α-GalCer. After 3 days, proliferation was assessed by [3H] thymidine incorporation (C), and culture supernatants were evaluated for IL-2 release by ELISA (D). Average + SEM is reported. p < 0.001 by two-way ANOVA and the Bonferroni multiple comparison test. *p < 0.05, ***p < 0.001.