Fig. 5. Phenotypic and functional profiling of IFN–CSC immunogenicity.
a, Flow cytometry analysis of proliferation rate of CFSE-stained CD8+ OT-1 T cells stimulated with PAR or CD44H OVA-expressing cells. The histograms represent the FC (mean ± s.e.m. and individual data points, n = 3 independent experiments) of nonproliferating CFSE+highCD8+ cells. b, Flow cytometry analysis of CD45− OVA-expressing PAR and CD44H cell resistance to CD8+ OT-1-mediated killing. The histograms represent the FC (mean ± s.e.m. and individual data points, n = 3 independent experiments) of dying PI+CD45− cells. c, Multiparametric flow cytometry analysis of the indicated IC molecules in MCA205 or AT3 cells. Data are presented as mean ± s.e.m. and individual data points, with number of biologically independent samples collected over three independent experiments reported. d, Flow cytometry analysis of TIM3 in CD8+ tumor-infiltrating lymphocytes from MCA205-derived tumor grafts 15 days post in vivo treatment with PBS, DOX (2.9 mg kg–1), or CDDP (2.5 mg kg–1). Data are presented as mean ± s.e.m. and independent data points for 15 mice per group from three experimental replicates. e, Quantification of released chemokines in supernatants from MCA205 and AT3 cells by Luminex Multiplex Assay. One representative experiment out of two is shown. f–i, Time-lapse analysis of H-2Kb splenocyte migration towards PAR and CD24L AT3 cells in microfluidic devices. Plots in (f) represent individual splenocyte trajectories towards target cancer cells (black spots) upon time-lapse recording. Quantification of interaction times between individual splenocytes and PAR or CD24L ICD–CSCs are shown in g, see also Supplementary Videos 1–4. Pictures of splenocytes in competition microfluidic devices (scale bar, 100 μm) and quantification of splenocytes migrated towards PAR or CD24L ICD–CSCs are shown in h and i. Data are expressed as mean ± s.e.m. and individual data points; number of biologically independent samples collected over three (f,g) and two (h,i) independent experiments is reported. See also Extended Data Fig. 4. a–c, Unpaired two-sided Student’s t-test and unpaired two-sided Student’s t-test followed by Welch’s correction. d,f,g, Two-tailed Mann–Whitney test compared with PBS (d) and CTR (f,g). i, Ordinary two-way RM ANOVA test followed by Bonferroni’s correction.