Extended Data Fig. 3: Tumour-derived PGE2 does not impact on priming of anticancer CD8+ T cells in lymph nodes.
From: PGE2 limits effector expansion of tumour-infiltrating stem-like CD8+ T cells
a, Experimental design for b-e. WT mice received 1 × 103 naive OVA-specific CD45.1+ OT-I T cells followed by inoculation with 2 × 106 control or Ptgs1/Ptgs2−/− BRAFV600E-OVA cells. 6 days later, tdLNs were analysed by flow cytometry. Mice injected with Ptgs1/Ptgs2−/− BRAFV600E cells (lacking OVA expression) served as control. b, Representative plot showing S8:H-2Kb surface staining on migratory cDC1 (identified as live CD45+CD11c+MHCIIhiCD103+CD8α−CD11b− cells). c, Quantification based on b, with n = 5 for control and Ptgs1/Ptgs2−/− BRAFV600E-OVA, n = 4 for Ptgs1/Ptgs2−/−BRAFV600E. d, Flow cytometry plots showing the sorting strategy for naive OT-I T cells. e, Flow cytometry plots showing expression of CD44 and TCF1 in polyclonal CD8+ T cells and OVA-specific OT-I T cells. f, Quantification of antigen-experienced CD44+ TCF1+ OT-I T cells based on e. n = 5. g, PGE2 concentration in lysates from tumours and indicated organs analysed 11 days after s.c. inoculation of WT mice with control or Ptgs1/Ptgs2−/− BRAFV600E melanoma cells (n = 5 per group). h, Effect of equilateral co-transplantation of 2 × 105 control and Ptgs1/Ptgs2−/− BRAFV600E tumours on tumour growth (n = 4 per group). Data in c and f-h are pooled from two (c,f,h) or three (g) independent experiments and depicted as mean ± s.e.m. Plots in b,e show data for one sample representative for n = 5 samples from two independent experiments. P values in c,f are from one-way ANOVA with Tukey’s multiple-comparison test, P values in g are from unpaired t-tests, P values in h are from two-way ANOVA with Bonferroni’s multiple-comparison test. P ≥ 0.05, not significant (NS).
