Fig 2: Preliminary clinical connections of mouse findings based on NCI human MM RNA-seq and germline mutation data (16). A, Comparison of CCL2 mRNA expression levels based on BAP1 germline mutation status in 100 MM samples from the NCI cohort. Statistical analysis was performed using the Student t test. B, Immunoblot analyses of CCL2 in human MM cell lines with or without loss of BAP1 expression. GAPDH was used as a loading control. C, Scatter plots depicting the relationship between CD163 and CCR2, ENTPD1 (CD39), IL10, IL10RA, MRC1 (CD206), and STAT3 mRNA expression levels. Statistical analysis for this correlation study was conducted using Spearman rank-order correlations (r) and P values.

Fig 3: Expression of IL6 and IL10 in ascites and Bap1 and p16Ink4a in MM tumors from chrysotile-exposed Bap1+/− mice. A, Concentration of IL6 in ascites of Bap1+/− mice compared with that from Bap1+/+ mice with MMs induced by chrysotile. Ascitic fluids were obtained from 5 Bap+/+ mice and 4 Bap1+/− mice with chrysotile-induced MMs. B, Expression of IL10 in ascites of 4 Bap1+/− mice versus that in ascites of 5 Bap1+/+ mice with MMs induced by chrysotile. C, Immunoblot analysis of tumor suppressors Bap1, Nf2, and p16Ink4a in MM tumors from 1 chrysotile-exposed Bap1+/+ mouse and 6 chrysotile-exposed Bap1+/− mice. NMC, normal (mouse) mesothelial cells; Vinculin, loading control.

Fig 4: MIIP modulates the STING–NFκB2–IL10 signaling pathway in CRC cells. Immunofluorescence staining showing the effects of MIIP knockdown and MIIP overexpression on dsDNA levels (A) and the distribution and expression of STING (B) in SW480 and SW620 cells. (C) Western blots showing the effects of MIIP knockdown and overexpression on STING, TRAF3, p-p100, and p52 levels. (D) ELISA results showing the effects of MIIP knockdown and overexpression on IL10 levels in the cell culture medium. (E and F) Effects of MIIP knockdown and/or STING knockdown on p52 expression (E) and IL10 levels in the cell culture medium (F). All data are presented as the means ± SDs (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001; scale bar, 50 μm.

Fig 5: Tlr4 expression in Tregs mitigates liver I/R injury in Treg-Tlr4–/– mice.(A) Schematic diagram of the adoptive transfer experiment. Treg-Tlr4–/– mice were administrated with PBS, Tlr4–/– Tregs, or WT Tregs via tail vein injection 12 hours before I/R. (B) Serum ALT and AST levels in the 3 groups of mice before and after I/R (n = 5 per group). (C) Representative H&E staining images of ischemia liver lobes in the 3 groups of mice before and after I/R (n = 5 per group). Scale bars: 200 μm. (D) Quantification of hepatic infiltrating CD11b+Ly6G+ neutrophils and CD11b+Ly6C+ monocytes by flow cytometry in the 3 groups before and after I/R (n = 5 per group). (E) Quantitative PCR analysis of Il1b, Tnf, and Il10 mRNA expression in ischemia liver lobes in the 3 groups of mice before and after I/R (n = 5 per group). (F) Serum IL-1β, TNF-α, and IL-10 levels in the 3 groups of mice before and after I/R (n = 5 per group). Statistical analyses were performed using 1-way ANOVA with Tukey’s post test (B–F). *P < 0.05, **P < 0.01, ***P < 0.001. ALT, Alanine aminotransferase; AST, Aspartate aminotransferase; I/R, ischemia/reperfusion; Tregs, regulatory T cells.