Fig. 5: DNAse I-treated mice show increased intratumoral CD8 infiltration post-RT.
From: Role of neutrophil extracellular traps in radiation resistance of invasive bladder cancer
a FFPE tumors were collected from C57BL/6 mice injected subcutaneously (s.c) with MB49 tumors from groups: non-irradiated control (Control), C57BL/6 mice treated with DNAse-treated mice (DNAse I), irradiated C57BL/6 (RT) and irradiated C57BL/6 treated with DNAse I (RT + DNAse I) at endpoint (3 weeks post-RT). Tissues were stained for immunofluorescence analyses with CD8 (red), NE (yellow), H3Cit (green), nuclei (blue). Representative confocal images from three independent experiments obtained with 20× objective, scale bar 50 μm. b Quantification of intratumoral CD8 T-cell infiltration using QuPathV6. Data represented as percentage CD8 T-cells per total cells per field (n = 7 mice per arm). c Schematic of tumor and survival experiment in athymic C57BL/6 mice. d Tumor growth kinetics of irradiated athymic mice injected s.c with MB49, treated with (n = 8 mice per group) or without DNAse I (n = 9 mice per group). e Kaplan–Meier percent survival. f Chromogenic immunohistochemical staining of athymic tumors, stained for PMNs and NETs (NE-blue, H3Cit-purple), scale bar 50 μm. g Quantification of PMN infiltration and NETs in tumors, data represented as percent positive of total cells per field of view (n = 5 mice per arm). Data represented as mean ± SEM, unpaired two tailed student’s t-test was used to assess statistical significance in (b, g) two way ANOVA with Bonferroni’s multiple comparison’s test (d), log rank (Mantel-cox) test (e). NS not significant (p > 0.05), *p < 0.05, **p < 0.01, ***p < 0.001.
