Fig 2: Lkb1 discriminates regulatory from inflammatory transcriptional programmes. a GSEA of transcriptional profiles of DCs from mice treated with LPS compared with that of Cd11cCreLkb1f/f mice. The upper part shows the list of gene sets related to inflammation; the lower part shows a representative plot of the gene set; –, no enrichment. b Scatter plot of Log2 counts of the genes expressed in splenic DCs from Lkb1f/f mice treated with LPS (n = 1) compared with those from Cd11cCreLkb1f/f mice (n = 3). Grey dots mark genes with less than two-fold change in any group. Red and blue dots mark the positively correlated genes with more than two-fold change from the two groups, while negatively correlated genes are indicated in green. The names of pro-inflammatory genes only upregulated in LPS-treated DCs are indicated in blue characters, and Ox40l, upregulated in both groups, is indicated in red characters. c mRNA levels of Cxcl11, Il1a, Il27, Cxcl9, Il15, Cd80, Il1f9, Cxcl10 and Ccl5 in splenic DCs from Lkb1f/f mice, Lkb1f/f mice treated with LPS, and Cd11cCreLkb1f/f mice. d mRNA level of Ox40l in DCs sorted from mice treated with or without LPS. e, f Flow cytometric analysis (e) and quantification of relative MFI (f) of OX40L protein expression on splenic DCs from mice treated with or without LPS (1.5 mg kg−1). The results are presented as the mean ± S.E.M., *P < 0.05, **P < 0.01, ***P < 0.001, by Student's t-test (c, d, f). Data are pooled from c, d, f or are representative of e at least two independent experiments with similar results

Fig 3: PQ/PB-Gel for triggering antitumor innate and adaptive immune response after B16F10 melanoma surgery. (A and B) Representative flow cytometric images (left) and relative quantitative images (right) of M2-like TAMs (CD206hi) and M1-like TAMs (CD80hi) gating on F4/80+CD11b+CD45+ cells. Data are presented as means ± SD (n = 5). (C) Representative flow cytometric images (left) and relative quantification images (right) of CD80+CD86hi DCs gating on CD11c+ cells. Data are presented as means ± SD (n = 5). (D) Relative quantitative images of CD11c+ cells. Data are presented as means ± SD (n = 5). (E) Relative quantification of CD3+ cells within the tumor. Data are presented as means ± SD (n = 5). (F and G) The quantification results of CD4+, CD8+ cells within the tumor in different groups. Data are presented as means ± SD (n = 5). (H) Representative flow cytometric analysis of T cells infiltration within the tumor gating on CD3+ CD45+ cells. (I) The ratio of CD8 to CD4 cells within the tumor in different groups. Data are presented as means ± SD (n = 5). *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 by one-way analysis of variance.

Fig 4: In vitro adhesiveness, cellular uptake, cytotoxicity, and ICD of evaluation of PexD. A Confocal microscope images of B16-F10 cells incubated with free DOX and PexD for 0.5 and 2 h, respectively. Scale bar: 10 μm. B Flow cytometry measurement of B16-F10 cells incubated with free DOX and PexD for 0.5 and 2 h. Scale bar: 10 µm. C Fluorescence intensity analyzed by flow cytometry. The data are expressed as mean ± SD (n = 3). D The adhesion of DiR-labeled Pex to B16-F10 cells with confocal laser scanning microscope. CD44 staining (green), DIR-labeled Pex (red) and nucleus (blue). Scale bar (upside): 10 µm. Scale bar (down): 5 µm. E Relative HMGB1 release from B16-F10 cells treated for 24 h with PexD or DOX (3 μg mL−1). The data are expressed as mean ± SD (n = 3). F Relative ATP release from B16-F10 cells treated for 24 h with PexD or DOX (3 μg mL−1). The data are expressed as mean ± SD (n = 3). G Representative flow cytometric analysis of CD80+ CD86+ cells. The data are expressed as mean ± SD (n = 3). C–F **P < 0.01; ***P < 0.001; ****P < 0.0001

Fig 5: Combining CXCR4-targeted p53 mRNA NPs with PD-1 blockade reprograms the immune TME and promotes antitumor immunity in ectopic HCC.a Bioluminescence images of the luciferase-expressing RIL-175 tumors grafted subcutaneously in C57Bl/6 mice after 6, 12, and 18 days of treatment (n = 3 mice/group). b Tumor growth rate in each treatment group (n = 7 mice/group; ***P < 0.001). c Western blotting analysis on the expression levels of p53 protein in the s.c. RIL-175 tumors after treatment. GAPDH was used as the loading control. d–f Flow cytometry analysis (n = 3 tumor samples from each group) of lymph node CD80+CD86+ dendritic cells gating on CD11c+ cells (d), and tumor-infiltrating CD8+CD3+ T cells (e) and M2-like CD206+F4/80+CD11b+ macrophages (f). g Representative immunofluorescence for CD8 (in red) to confirm intratumoral T cell infiltration after treatment with CTCE-EGFP NPs, anti-PD-1 (aPD1), CTCE-p53 NPs, or the combination. Scale bar: 200 µm. h–k Protein array analysis of differential expression of cytokines in s.c. HCC tissues after treatment (n = 3 samples per group): TNF-α (h), IL-1β (i), IFN-γ (j), and IL-6 (k). Statistical significance was calculated using one-way ANOVA with a Tukey post-hoc test. All data are presented as mean ± S.D. For c and g: this experiment was repeated thrice independently with similar results. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Source data are provided as a Source Data file.