Fig. 5. IMD_1-53 decreased the CHOP level and apoptosis in vivo and in vitro.

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 intermedin_1-53 (IMD_1-53) during the left 6 weeks of high-fat diet feeding. Representative images and quantification data of CHOP (A) and cleaved caspase-3 (B) immunohistochemical staining, and terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) staining (C) at the aortic root of mice from con, HF, and HF + IMD. Black arrows indicate the area stained positively for CHOP or cleaved caspase-3. Scale bars, 200 μm (A, B) and 100 μm (C). n = 6. Data are mean ± SD. ^*P < 0.05, ^**P < 0.01 compared with Con, ^#P < 0.05 compared with HF group; one-way ANOVA. D, E Western blot analysis of protein expression of CHOP (D) and cleaved-caspase3 (E) in macrophages treated with PBS, IMD_1-53, ox-LDL, and IMD_1-53 + ox-LDL. β-actin was a control for protein loading. Results are representative of four experiments. Densitometric analysis of protein levels is shown as a ratio to β-actin. n = 4. Data are mean ± SD. ^*P < 0.05, ^**P < 0.01 compared with con, ^#P < 0.05, ^##P < 0.01 compared with the ox-LDL group; one-way ANOVA. F Representative images and quantification data of Hoechst staining in macrophages treated with PBS, IMD_1-53, ox-LDL and IMD_1-53 + ox-LDL. Scale bars, 100 μm. n = 6. Data are mean ± SD. ^**P < 0.01 compared with con, ^##P < 0.01 compared with ox-LDL group; one-way ANOVA.