Fig 1: METTL3 upregulates NSD2 to alleviate interstitial fibrosis in the kidney tissues of mice with DN. A Kidney/body weight of mice; B Levels of FBG, SCr, S-Cys-C, 24-h U-protein, U-Cys-C, and SBP in mice; C Levels of SOD, MDA, IL-6, MCP-1, and hydroxyproline in mouse kidney tissues determined by ELISA kits; D Protein levels of METTL3, NSD2, and the interstitial fibrosis-related proteins COL1A1, Fibronectin and E-cadherin in mouse kidney tissues determined by western blot analysis; E Pathological changes in mouse kidney tissues determined by HE staining; F Collagen deposition in mouse kidney tissues determined by Masson’s trichrome staining. There were at least five mice in each group. Data were collected from three independent experiments and expressed as the mean ± SD. Differences were analyzed by two-way ANOVA (A, B, C, D and F). *p < 0.05
Fig 2: 4-octyl itaconate inhibits the inflammatory reaction in mouse ATDC5 cells and human C28I2 cells. The ATDC5 cells and C28I2 cells are induced with LPS (10 µg/ml) for 12 h and then treated with OI (100 µm) for 12 h. The protein levels of Il-6, MCP-1, Il-10 and Tnf-a are detected by ELISA (A–D, H, and I). The mRNA in cells is detected by qPCR (E, F, J and K). The NF-?B p65 transcriptional activity in nucleus is detected by the NF-?B p65 Transcription Factor Assay Kit (G and L). Each experiment is repeated three times. Data are presented as mean ± standard deviation (n = 3). Symbols ‘*’and ‘#’ indicate the significant difference set at p < 0.05 in comparison with control (‘Ctrl’) and LPS induction respectively
Fig 3: AN1284 ameliorates hepatic steatosis and inflammation in db/db mice. The db/db mice exhibit increased liver weight (A), serum ALT (B), and ALP (C) levels as well as hepatic triglyceride and cholesterol contents (D,E). These parameters were prevented or ameliorated by AN1284 treatment. AN1284-treated mice also displayed decreased serum triglyceride levels (F), increased HDL/LDL cholesterol ratio (G), without an effect on total cholesterol levels (H). Quantification of hepatic fat content revealed an increase in db/db animals, which was normalized by AN1284 treatment (I). Representative liver images demonstrating macrovesicular steatosis in H&E-stained sections from db/db mice. Scale bar, 100 µm (J). The elevated hepatic mRNA expression levels of Acc (K), Scd1 (L), and Cd36 (M) in db/db mice were normalized by AN1284 treatment. Insignificant changes in hepatic TNFa protein levels were noted in non-treated and treated mice (N). Reduced protein expression of IL-18 (O), and MCP1 (P) in AN1284-treated db/db mice was documented. Scale bar, 50 µm. M1-to-M2 macrophage marker ratios displayed high content of M1 macrophages in db/db animals, which was diminished by AN1284 treatment (Q,R). Data represent the mean ± SEM from 8 to 10 mice per group. *P < 0.05 relative to control non-diabetic mice, #P < 0.05 relative to db/db-Veh-treated control mice.
Fig 4: Loss of Cideb protects mice from diet‐induced fatty liver diseases ALiver morphology of WT and Cideb −/− liver under chow diet and HFLF diet. H/E staining, oil red‐O staining, immunohistochemistry, and ultrastructure (Electron Microscope) were performed. Scale bar represented 50 μm in the upper three rows of images and 2 μm in the bottom row of images. LD: lipid droplet; ER: endoplasmic reticulum; M: mitochondrial; N: nucleus.B, CTotal liver TAG levels (B) and liver cholesterol ester levels (C) of WT and Cideb −/− mice under chow diet and HFLF diet (n = 8 per group).DLoss of Cideb inhibits diet‐induced SREBP activation. IB of hepatic SREBP processing and lipogenic enzyme levels (Fasn, Scd1) of WT and Cideb −/− mice under chow diet and HFLF diet.ESerum AST and ALT levels of WT and Cideb −/− mice under chow diet and HFLF diet (n = 8 per group).F Cideb deficiency alleviates the whole body inflammation response. Serum TNFα, MCP1, and IL6 levels of WT and Cideb −/− mice under chow diet and HFLF diet (n = 8 per group).G, HSerum fed glucose levels (G) and insulin levels (H) of WT and Cideb −/− mice under chow diet and HFLF diet (n = 8 per group).IGlucose tolerance test (GTT) of WT and Cideb −/− mice under HFLF diet (n = 5 per group).JInsulin tolerance test (ITT) of WT and Cideb −/− mice under HFLF diet (n = 5 per group).Data information: Data represent Mean ± SEM; NS: not significant; *P < 0.05; **P < 0.01; ***P < 0.001, by 2‐tailed Student's t‐test.Source data are available online for this figure.
Fig 5: AN1284 prevents the renal inflammatory response in type 2 diabetic mice. The elevated renal protein expression levels of the inflammatory markers MCP-1 (A,E), TNFα (B,F), TGFβ (C,G), and IL-18 (D,H) in db/db mice were normalized by AN1284 treatment. Scale bar, 50 μm. Data represent the mean ± SEM from 8 to 10 mice per group. *P < 0.05 relative to control non-diabetic mice, #P < 0.05 relative to db/db-Veh-treated control mice.
Supplier Page from Abcam for Mouse MCP1 ELISA Kit