Fig 2: Hepatic inflammatory responses correlate with impaired infant growth and development. The mice were exposed to a liquid ethanol diet for 2 weeks and simultaneously, 4-MP (10 mg/kg, i.p., n = 8/group) was administered three times per week for 2 weeks. (a) Serum proinflammatory cytokines for IL-6, TNFα, MCP-1, and IL-1β. *p < 0.05 vs. pair-fed mice with saline, **p < 0.01 (n = 8 dams/group). (b) Real-time PCR analyses of hepatic mRNA expression of proinflammatory cytokines in the livers of E15.5 mice. (c) Quantitative mRNA analysis of CCR2 and F4/80 in the livers of E15.5 mice. (b,c) *p < 0.05 vs. pair-fed mice with saline, **p < 0.01 (n = 8 dams/group). (d) Representative images of liver sections stained with antibodies against CD14 for monocytes and against F4/80 for macrophages, analyzed by confocal microscopy (scale bars, 10μm) and quantified data. *p < 0.05 vs. pair-fed mice with saline, **p < 0.01 (n = 8 dams/group). (e) Representative Western blotting (left) and quantified data (right) for NF-κB activation and TLR4 in livers of E15.5 mice. *p < 0.05 vs. pair-fed mice with saline, **p < 0.01 (n = 8 dams/group). (f,g) Correlation between TNF-α mRNA levels and postnatal body weight (P0 macrosomia, f) and P21 growth retardation (g), respectively. (h and i) Correlation between TLR4 mRNA levels and P0 macrosomia (h) or E15.5 lipid accumulation (i). Correlation coefficients (r) and p values are given. Data are expressed as the mean ± SEM. Data in a-d were analyzed by two-way ANOVA with the post hoc Bonferroni’s multiple comparison test. Data in e were analyzed by one-way ANOVA with Tukey’s post hoc multiple comparison test. Data in f-i were performed by correlation analyses using Pearson’s analysis.

Fig 3: EXOKV11 has therapeutic effect in post-treatment model. (A) Schematic depiction of the post-treatment procedure in VEGF-induced vascular leakage model. 100 ng VEGF was intravitreally injected to induce vascular leakage in the retina. After 24 h, EXOKV11, KV11, EXO, or vehicle was retro-orbitally injected in adult wild type C57BL/6J mice. The mice were sacrificed for analysis after another 48 h. (B) FITC-dextran was injected and the retinas were harvested after treatment. Representative images of flat-mounted retina show extravasated FITC-dextran and CD105+ vessels. (C) Quantification of dextran leakage in (B) (n = 4 - 6 mice per condition). (D) Representative images of F4/80+ macrophages (green) and CD105+ vessels in retinas treated as in (A). (E) Quantification of macrophage infiltration in (D) (n = 4 - 6 mice per condition). The data represent as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA followed by Tukey's multiple comparisons test in (C), (E). Scale bars, 50 µm.

Fig 4: Myeloid AT1R gene deficiency reduced macrophage infiltration and vascular inflammation in DOCA/salt hypertensive mice. Representative images of aortic F4/80 expression assessed by immunofluorescence staining (A); quantitative analysis of average immunofluorescence intensity of F4/80 expression (B); protein expressions of proinflammatory cytokines, TNFα (C) and IL-1β (D), in the aorta; protein expression of eNOS (E) and p-eNOS [Ser 1177, (F)] in the aorta. Statistical comparisons were carried out with two-way ANOVA followed by the Tukey post hoc test. Data were expressed as mean ± SEM. N = 8, *p < 0.05 vs. control mice. # p < 0.05 vs. DOCA group.

Fig 5: Significantly different expressions of IL1A (A), IL1B (B), and ESR1 (C) from the LLC mouse model in different groups of mice treated with immune checkpoint inhibitors. The markers of macrophage (F4/80) and classic immune checkpoint (PD-L1) inhibitor were expressed differently among the border, the center of the subcutaneous model, and the orthotopic model of LLC (D).