Fig. 6: DTIC/Epacasome-2 fortified the PD-1 blockade to elicit tumour regression and complete metastasis remission in late-stage metastatic melanoma model.

a Drug administration timeline scheme in s.c. B16–F10-Luc2 tumour mice (n = 6 mice for (a–g), tumours: ~300 mm³). Mice were intravenously injected with DTIC, Epacasome-2 at eq. 75 mg DTIC/kg (based on the maximum tolerated dose (MTD) of DTIC)⁵¹ or 41 mg EPA/kg on day 12, 14 and 16 alone or in combination with i.p. α-PD-1 (100 µg per mouse) on days 12 and 15. b Average tumour growth curves measured by a digital calliper. c Representative mice Lago bioluminescence imaging (BLI) on day 12, 15 and 18. d Representative ex vivo BLI for various organs on day 18. e Tumour metastatic rate summarised in a heatmap. f Normalised BLI in various organs. g Intratumoural IL-2 level. h–m Representative flow histogram and quantitative analysis for intratumoural Rae-1 (h, i), Mult-1 (j, k), and MHC-I (l, m) on tumour cells (CD45^- cells). n Representative flow cytometric plots of intratumoural NKG2D⁺, CD69⁺, IFN-γ⁺, Perforin⁺ and Granzyme B⁺ associated NK⁺ cells and their respective quantification (right panel, right panel, for (h–n), n = 3 tumours were randomly chosen from an independent study on day 18). The flow cytometric gating strategies are placed in Supplementary Figs. 20 and 21. Intratumoural NKG2D⁺, CD69⁺, IFN-γ⁺, Perforin⁺, Granzyme B⁺, PD-1⁺, Lag-3⁺ and Tim-3⁺ associated CD8⁺ T cells flow cytometric data and gating strategies are placed in Supplementary Figs. 19 and 22. Data in (b, f, g, i, k, m, n, right panel) are expressed as mean ± s.d. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons. Source data are provided as a Source Data file.