Fig 1: IMD1-53 inhibited apoptosis-triggered inflammasome in advanced lesional macrophage.A–C Eight-week-old male ApoE-/- were fed a standard chow diet (con) or a high-fat diet (HFD) for 16 weeks. After 10 weeks of HFD feeding, ApoE–/– mice received either PBS or intermedin1-53 (IMD1-53) during the left 6 weeks of high-fat diet feeding. Quantitative real-time PCR of interleukin6 (IL6), tumor necrosis factor a (Tnfa), and Nlrp3 mRNA expression (A). Results are relative to the GAPDH level (n = 3). Representative images and quantification data of apoptosis-associated speck-like protein containing CARD (ASC) (B) and IL-1ß (C) immunohistochemical staining at the aortic root of mice from con, HF, and HF + IMD. Black arrows indicate the area stained positively for ASC or IL-1ß. Scale bars, 200 µm. n = 6. D Plasma IL-1ß level was measured. n = 6. Data are mean ± SD. *P < 0.05 and **P < 0.01 compared with con, #P < 0.05 and ##P < 0.01 compared with HF group; one-way ANOVA. E Western blot analysis of protein expression of IL-1ß in macrophages treated with PBS, IMD1-53, ox-LDL and IMD1-53 + ox-LDL. ß-actin was a control for protein loading. Results are representative of four experiments. Densitometric analysis of protein levels is shown as a ratio to ß-actin. n = 4. F IL-1ß in the cell culture supernatant was also measured. n = 5. Data are mean ± SD. **P < 0.01 compared with con, #P < 0.05 compared with the ox-LDL group; one-way ANOVA.
Fig 2: Effect of DJ-1 overexpression on expression of Nrf2-NLRP3-axis associated proteins in hippocampus of 5XFAD transgenic mice. (A) Western blotting and quantification of cytoplasmic and nuclear Nrf2 protein. β-actin and histone H3 were used for normalization for cytoplasmic and nuclear proteins, respectively. (B) Western blotting and quantification of NLRP3 and ASC protein expression. n=10. Data are presented as the mean ± standard deviation. *P<0.05, **P<0.001 vs. sham group. Nrf2, nuclear factor erythroid 2-related factor 2; NLRP3, NLR pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD; NC, negative control.
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