Fig 2: Serum TNC can reflect the immune microenvironment of LGGs and predict the effect of immunotherapy. (A) qRT-PCR analysis of mRNA expression of TNC in normal brain tissues (n = 6), grade II (n = 15) and grade III (n = 17) glioma patients. (* p < 0.05; ** p < 0.01) (B) Scatter plot displaying the serum concentration of TNC in normal brain tissues (n = 6), grade II (n = 15) and grade III (n = 17) glioma patients. (** p < 0.01; *** p < 0.001) (C) Scatter plot demonstrating the correction between TNC serum concentration and immunity-status in grade II and III glioma patients (Grouped based on the median of the tumor CD11b mRNA level. ***p < 0.001; n = 32) (D, E) ROC curves showing the diagnostic value of serum TNC level for distinguishing the status of LGG immune microenvironment. (F, G) TIDE scores and T cell exclusion scores in different TNC mRNA expression subgroups. The score between the two subgroups were compared through the Wilcoxon test. (Top: TNC high expression; Bottom: TNC low expression. * p < 0.05, *** p < 0.001) (H) The potential application of TNC in the early diagnosis and individualized treatment of LGG patients.

Fig 3: Correlation between DHCR7 expression and macrophage polarization in MASH. (a) Relative expression of cholesterol biosynthesis and efflux genes in primary macrophages after ox-LDL (100 µg/ mL) or ox-LDL combined with statin treating for 24 h. (b) Expression levels of cholesterol biosynthesis and efflux genes in primary macrophages polarized to M1 with LPS (100 ng/mL) or M2 with IL-4 (20 ng/mL) for 24 h. (c-d) Western blot analysis for DHCR7 protein in M1 and M2 macrophages, with densitometry. (e) Immunofluorescence staining for DHCR7 in the polarized-macrophages, 200× magnification. (f-g) Multiple immunofluorescence co-localization for DHCR7 and CD11B on livers of NCD or CDAHFD mice. (h-i) DHCR7 protein and mRNA levels in total liver and primary macrophages isolated from livers of mice on control and CDAHFD. (j-k) Immunofluorescence multi-labeling of liver sections from NC and MASH patients for DHCR7 and macrophage marker CD68 to determine the proportion of DHCR7+CD68+ cells among total CD68+ cells. Data are presented as mean ± SD, with n = 3 per group. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 between groups

Fig 4: MC-specific Atg16l1 deficiency increased neutrophil infiltration in the colonic mucosa and tumor of ApcMin/+ mice upon CoPEC infection dependently of colibactin. ApcMin/+/Atg16l1[ΔMC] and ApcMin/+ mice were orally administered with PBS or with 109 11G5 or 11G5∆clbQ bacteria, and sacrificed at day 65 post-administration. Representative images of LY6G (red) and ITGAM (green) immunofluorescent double staining in the colonic mucosa (A) and tumors (B). Nuclei were stained with Hoechst (blue). Bars: 100 μm. Boxes show magnified fields. Quantification of LY6G+ ITGAM+ neutrophil number/mm2 of colonic mucosa (C) or tumor (D). Statistical analysis was performed using one-way Anova test followed by Bonferroni posttest. **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001.

Fig 5: Myeloid-macrophage-specific DHCR7 deficiency exacerbates MASH progression. Wildtype (WT) and myeloid-specific DHCR7 knockout (DHCR7 cKO) mice were fed with a normal control diet (CON) or a CDAHFD for 10 weeks (n = 7). (a) Body weight progression, (b) liver/body weight ratio, and (c-d) serum ALT and AST levels were detected. (e-f) Hepatic TG and TC levels were measured. (g) ELISA quantification of liver IL-1β and IL-6 levels. (h) Histological analysis of liver sections is presented through H&E, ORO staining, and immunohistochemistry for F4/80+ and CD11b+ cells (200× magnification). (i) NAS score. (j-k) Relative protein levels of CD86, ARG-1, CD206 were determined, with densitometry. (l) Flow cytometry of primary macrophage in cKO mice and WT with different diets. (m) Flow cytometry of TREM2 in macrophages. (n) Predominantly of the M1 type in macrophages, including quantification of both the percentage of M1 macrophages and M1 macrophage counts per liver weight. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 between group