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: TNC was related to GAM cell infiltration in LGGs (A) Kaplan-Meier survival analysis showing that the infiltration level of macrophages, M1-like cells, and M2-like cells with the prognosis of LGG patients in the TIMER database using the XCELL method. (** p < 0.01; *** p < 0.001; **** p < 0.0001) (B) The scatter plot indicating the correlation between TNC expression level (log2TPM) and the infiltration level of macrophages, M1-like cells, and M2-like cells in LGG patients using the TIMER database. (*** p<0.001) (C) qRT-PCR analysis of Iba1 and CD11b mRNA expression in 32 patients grouped by TNC mRNA level. (* p < 0.05; *** p < 0.001) (D) IF staining of TNC (green) and Iba1 (red) in human LGG samples. Representative images are shown. (Scale bars = 200 um) (s) MRI images of patients with grade II (E) and grade III (F) gliomas.(n=16) (G) Immunoblotting of TNC and CD11b in grade II and III glioma patients sorted by CD11b mRNA level. (The samples correspond to the patients in (E, F).

Fig 4: Hepatic cholesterol accumulation and M1 macrophages infiltration in MASH. Six-week-old male C57BL/6J mice were fed either a NCD or a CDAHFD for six weeks (n = 3). (a) Body weight gain curves and (b) liver/body weight ratio was recorded. (c) Serum ALT and AST levels were detected. (d) Liver sections stained with H&E and ORO, and immunohistochemically for F4/80+ and CD11b+, 100× magnification. (e-f) Quantifies hepatic TC and TG levels. (g) NAS score. (h-j) Hepatic relative mRNA expression of Tnfa, Il1b and Il6. (k-l) Hepatic relative protein levels of CD86, ARG-1 and CD206. (m-o) Flow cytometry analysis of macrophages (live+CD45+F4/80+CD11b+) and M1 macrophages (live+CD45+F4/80+CD11b+iNOS+). (p-q) Flow cytometry analysis of TREM2 (live+CD45+F4/80+TREM2+). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 between groups

Fig 5: 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