Fig 1: MiR-21-3p inhibits adipose browning to promote HG-induced myocardial fibrosis. Preadipocytes from miR-21 KO mice were induced to differentiate into mature adipocytes and then transfected with mimics NC, miR-21-3p mimics, or miR-21-5p mimics. Mature adipocyte coculture with atrial fibroblasts from WT mice treated with LG (5.5 mM) and HG (30 mM) for 72 h. (a) TGF-β1, CTGF, collagen I, and collagen III expressions in atrial fibroblasts were analyzed by western blot and quantified by Scion Image software. (b) Representative images and quantification of collagen I and collagen III expressions in atrial fibroblasts were analyzed by immunohistochemistry, scale bar = 100 μm. (c) ELISA was used to analyze IL-6, TNF-α, and MCP-1 concentrations in coculture supernatants. (d) Western blot was used to analyze UCP1 expression in mature adipocytes and quantified by Scion Image software. (e) Representative images and quantification of UCP1 expression in mature adipocytes were detected by immunofluorescence, scale bar = 50 μm. UCP1 (red) was shown; nuclei were stained by DAPI (blue). The data are presented as the mean ± SD of three independent experiments. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.
Fig 2: Inflammation levels in brain tissues after clusterzyme treatment.a–c Western blotting for IL-1β, IL-6, and TNFα in the brain tissues 1, 3, and 7 days post TBI after treatment (n = 3 per group), respectively. d Western blotting quantitative analysis of inflammatory factors at different time points (n = 3 per group). All the samples were derived from the same experiment and blots were processed in parallel. Data are presented as mean ± SEM and compared with the Sham and TBI groups, analyzed by one-way ANOVA with two-sided LSD test (adjusted p values are shown). It can be seen that Au24Cd1 can rapidly and significantly reduce the upregulated inflammatory cytokines of IL-1β and IL-6 after brain injury, while Au24Cu1 has a better ability to reduce the expression of TNFα. e–g ELISA quantitative analysis of IL-1β, IL-6, and TNFα levels in brain tissues on days 1, 3, and 7 with or without clusterzymes treatment (n = 5 per group), respectively. Data are presented as mean ± SEM and compared with the Sham and TBI groups, analyzed by one-way ANOVA with two-sided LSD test (adjusted p values are shown). h Immunofluorescence co-staining of IL-1β and microglia (Iba-1), astrocytes (GFAP), or neurons (NeuN) in injured cortex 3 days post injury with or without clusterzyme treatment. Quantitative analysis of i the number of IL-1β+ expression in different positive cells and j the pixels density of Iba-1/NeuN/GFAP cells in the injured cortex with or without clusterzyme treatment (n = 3 per group). Data are presented as mean ± SEM and compared with the Sham and TBI groups, analyzed by one-way ANOVA with two-sided LSD test (adjusted p values are shown). Experiments were repeated independently a–c twice and h three times with similar results.
Fig 3: Nano-HO improved the spatial learning and memory functions of TgCRND8 mice. The spatial learning and memory functions were evaluated using MWMT (n = 9). (A) Experimental design and treatment schedule to evaluate the neuroprotective effects of HO and Nano-HO on TgCRND8 transgenic mice; (B) The latency to find a hidden platform during 4 consecutive days of training; (C) Time spent in the target quadrant; (D) Number of target crossings; (E) Representative swimming tracks of mice in the probe test. Nano-HO decreased the levels of cytokines including TNF-α (F), IL-1β (G) and IL-6 (H), enhanced the level of CCR2 (I) in the brain tissues of TgCRND8 mice (n = 6). Data were expressed as mean ± SEM. #p < 0.05 and ##p < 0.01 when compared with the WT group; * p < 0.05 and ** p < 0.01 when compared with the Tg + vehicle group; ▲ p < 0.05 when compared with free HO group.
Fig 4: Complement and cytokine assay and biochemical analysis of organ functions following multiple transfusions.(A) Timeline of the three succeeding blood transfusions. (B) Life span of PKH-26–labeled native and engineered RBCs after blood transfusion in vivo. (C) Serum levels of complement 3 and complement 4 after multiple transfusions. (D) Serum levels of TNF-α and IL-6 after multiple transfusions. (E) Biochemical analysis of liver function after multiple transfusions. ALT (U/l), AST (U/l), ALP (U/l), TBIL (μM), DBIL (μM). (F) Biochemical analysis of kidney function after multiple transfusions. CREA (μM), BUN (mM), UA (μM).
Fig 5: ADMA treatment exacerbates PM2.5-induced systemic inflammation, lung fibrosis, vessel remodeling and cell death. Mice were exposed to PM2.5 (10 mg/kg) with or without ADMA (2 mg/kg) every other day via intratracheal instillation for 2 weeks. A–D Serum TNFα (A), IL-6 (B) and ADMA C levels and mRNA levels of TGF-β and collagen I and III D were measured. E Representative lung sections were stained with H&E (the blue arrows point to widening of alveolar spaces, the green arrows point to inflammatory cell infiltration, the red arrows point to alveolar structure collapse), Masson’s trichrome kit (the black arrows point to collagen deposition), antibodies specific for αSMA (green) and CD31 (red), and TUNEL kit (red). Scale bar = 50 μm. F–H The relative collagenous fiber area (blue color area), muscularized vessels and TUNEL-positive cells (red point) were quantified. I Lung lysates were examined by Western blot. In Figure. A–D, N = 5; in Figure. F–H, N = 6; in Figure. I, N = 3; data are presented as the mean ± SD; * indicates p < 0.05, ** indicates p < 0.01
Supplier Page from Abcam for Mouse IL-6 ELISA Kit