Fig 2: Myeloid COX-2–/– mice had increased ATMs with impaired ability to phagocytose apoptotic adipocytes in DIO.WT and myeloid COX-2–/– mice were on the HFD for 12 weeks. (A) Myeloid COX-2–/– mice had increased Emr1 and Cd68 expression in EF and IF (n = 6). (B) Myeloid COX-2–/– mice had increased CLSs in EF, as indicated by CD68 staining (n = 8). Scale bar: 100 µm. (C) Myeloid COX-2–/– mice had increased proinflammatory mRNA and protein expression of TNF-a, IL-6, and IL-1ß in EF (n = 5 and 6). Scale bar: 200 µm. (D) ELISA determined higher EF TNF-a expression in myeloid COX-2–/– mice (n = 5). (E) Flow cytometry showed more proinflammatory ATMs (CD45+CD11b+F4/80+CD11c+) in myeloid COX-2–/– mice (n = 5). (F) EF ATMs isolated from myeloid COX-2–/– mice had increased proinflammatory cytokines (Tnf, Il6, and Il1b) but decreased expression of genes relating to lipid metabolism (Plin2, Cd36, and Gas6), indicating impaired ATM MMe polarization (n = 5). (G) Representative images showed that EF ATMs from myeloid COX-2–/– mice had decreased expression of LAMP2A, a marker of MMe polarization. Scale bars: 50 µm. (H) HMGB1 was localized to nuclei of adipocytes and stromal cells in WT mouse EF but was detected in nuclei and cytosol in myeloid COX-2–/– mice. Scale bars: 50 µm. (I) In vitro, the ability to phagocytose apoptotic adipocyte contents (green) was impaired in COX-2–/– PMs (PMs, F4/80 staining, red) isolated from myeloid COX-2–/– mice, as indicated by decreased lipid vesicles (asterisks) (n = 6). A, adipocytes; M, peritoneal macrophages. Scale bars: 50 µm. Data are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, analyzed using 2-way ANOVA followed by Bonferroni’s post hoc test for A and 2-tailed Student’s t test for B–F and I. EF, epididymal fat; IF inguinal fat.

Fig 3: Changes in IL-6 level. a IgAN group showing a significantly higher renal IL-6 level than the normal group based on western blot analysis (n = 7), b IgAN group showing a significantly higher serum IL-6 level than the normal group based on ELISA (n = 6). *p < 0.01, compared with the normal group, #p < 0.05, compared with the normal group. IgAN, IgA nephropathy; IL-6, interleukin 6.

Fig 4: IL-6 affects VE-cadherin expression in HRGEC cells via trans-signaling pathway. a–c Significantly decreased IL-6R protein and mRNA expression levels after lentivirus transfection, indicating successful IL-6R silencing. d After IL-6R silencing, IL-6/IL-6R still showed the ability to downregulate VE-cadherin expression via the trans-signaling pathway. e The sh-IL-6R/IL-6/IL-6R group showing significantly lower VE-cadherin expression than the control group (p < 0.01), but higher than the IL-6 group (p < 0.01). f After the use of sgp130Fc to block the trans-signaling pathway, VE-cadherin expression significantly increased. j The sgp130Fc/IL-6/IL-6R group showing significantly higher VE-cadherin expression than the IL-6/IL-6R group (p < 0.01) (n = 3). *p < 0.01, compared with the control group; #p < 0.01, compared with the IL-6/IL-6R group. IL-6, interleukin-6; IL-6R, interleukin-6 receptor; sh-IL-6R, IL-6R silenced; Lv-NC, negative control.

Fig 5: USMB-shMincle markedly inhibits M1/M2 transition of TAMs in vivo(A–D) USMB-shMincle dramatically increased the M1 phenotype (iNOS) but decreased M2 phenotype (CD206) and the protumoral effector IL-6 in TAMs. (E and F) USMB-shMincle inhibited the production of Mincle/Syk/NF-?B downstream effector IL-6, detected in the A549 xenograft by western blot and real-time PCR. Data represent mean ± SEM of 5 mice/group. Statistical analysis based on ordinary one-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 versus CTL; ####p < 0.0001, ###p < 0.001, ##p < 0.01, #p < 0.05 versus EV. Scale bars, 100 µm.