Fig 2: URB inhibited distant tumor growth and metastasis.a Schematic showing the treatment schedule in a distant tumor mouse model. b Primary tumor growth curve of mice treated with different treatments. n = 9 biologically independent animals per group (Asterisks: URB + US vs every other group). c Distant tumor growth curve of mice treated with different treatments (Asterisks: URB + US vs every other group). d Survival curves for different treatment groups. e Lung photographs and corresponding H&E-stained lung slices collected from the different group mice after treatment. f The number of metastasis foci in the lung of different groups (n = 5). g The weight of the lung of different groups (n = 5). h, i Flow cytometric analysis of M1 macrophages (CD80+F4/80+) in spleen. j, k Flow cytometric analysis of M2 macrophages (CD206+F4/80+) in spleen. l Flow cytometric quantification of CD4+ T cell (CD4+CD3+) and CD8+ T cell (CD8+CD3+) in spleen. m Flow cytometric quantification of Ki67+ in CD4+ and CD8+ T cells in spleen. n Flow cytometric quantification of Treg (FOXP3+CD4+) in spleen. o Flow cytometric quantification of central memory CD4+ and CD8+T cell in spleen. p, q Flow cytometric analysis and quantification of functional CD8+ T cells (TNF-α+CD8+) in spleen. r IFN-γ levels in serum at 1, 3, 5, 7 days after treatments. (Asterisks: URB + US group vs every other group). s TNF-α levels in serum at 1, 3, 5, 7 days after treatments (Asterisks: URB + US group vs every other group). t Flow cytometric quantification of effector memory CD4+ and CD8+T cell in distant tumor. u TGF-β levels in distant tumors at 1, 3, 5, 7 days after treatments (Asterisks: URB + US group vs every other group). For Fig. 6 (h–u), n = 4 biologically independent animals per group. Data were presented as mean ± S.D. Statistical analysis was calculated by using one-way analysis of variance with a Tukey’s test (****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05;). Source data are provided as a Source Data file.

Fig 3: Macrophage infiltration in LPS-induced noncanonical pyroptosis of acute keratitis in mice. Mice were sham-operated or intrastromal injected with LPS (3 µL 1 mg/mL) and treated with J114 or control four times a day. (A) Representative clinical images of LPS-induced acute keratitis in mouse corneas. (B) Mouse corneal clinical scoring was based on a 4‐point system. Corneal edema, opacity, and swelling was most obvious 48 hours after LPS injection (n = 12). (C) Representative histopathological examination (H&E staining) images of LPS-induced acute keratitis in mouse corneas. (D) Gating strategy for single cells, leukocytes, macrophages, and M1 phenotype macrophages. (E) The percentage of CD45+CD11b+F4/80+CD86+ cells in corneal tissue at 48 hours after LPS injection (n = 12). (F, G) Immunofluorescence staining displayed the colocalization of CD11b+ with IL-1β and F4/80 with IL-1β in the corneas after LPS intrastromal injection. (H, I) Immunohistochemical analysis was performed to examine the expression of macrophage CD11b+ and F4/80. These results were representative images of three independent experiments.

Fig 4: Concomitant TTFields with anti-PD-L1 reduced tumor volume and increased tumor infiltration of cytotoxic T cells. (A) Mice were orthotopically inoculated with LLC-2 Lewis lung carcinoma cells and treated according to the depicted timeline with: (1) isotype control, sham; (2) anti-PD-L1, sham; (3) isotype control, TTFields; or (4) anti-PD-L1, TTFields. (B) Tumor volumes were measured at treatment end. Tumors were further analyzed for the percentages of tumor leukocytes (CD45+) (C), macrophages (CD45+CD11b+F4/80+) (D), dendritic cells (CD45+CD11c+F4/80–) (E), myeloid-derived suppressor cells MDSCs (CD45+Gr-1+CD11b+) (F), as well as cytotoxic (CD3+CD8+) (G), helper (CD3+CD4+) (H), and regulatory (CD4+Foxp3+) (I) T cells, and the ratio of regulatory to cytotoxic T cells was calculated (J). MFI-median fluorescence intensity. Three independent experiments are shown. n = 14–17 mice per group. Values are mean ± SD. * p < 0.05, ** p < 0.01, and *** p < 0.001; one-way ANOVA followed by Tukey’s multiple comparisons for tumor volume; Student’s t-test for flow cytometry immunophenotyping.

Fig 5: In vivo immunogenicity. (A) Flow cytometric analysis of M1 macrophages (MCHII+CD80+) in spleen; (B) Flow cytometric analysis of M2 macrophages (F4/80+CD206+) in spleen; (C) Flow cytometric analysis of DCs (CD86+CD206+) in spleen; (D) Flow cytometric analysis of CD4+T-cell (CD4+CD3+) and CD8+T-cell (CD8+CD3+) in spleen.