Figure 4.

Effect of GYY4137 on NF-κB activation in RAW 264.7 cells. RAW 264.7 cells were exposed to ox-LDL (100 μg·mL⁻¹) in the presence or absence of GYY4137 (50–400 μmol·L⁻¹) or ZYJ1122 (200 μmol·L⁻¹) for 24 h. Western blots and quantification of NF-κB p65 protein expression (A). Representative examples of Western blots and quantification of IκBα phosphorylation (B), IκBα degradation (C) and ratio of IκBα phosphorylation to IκBα (D). NF-κB binding activity was evaluated by EMSA: (E-a) 1. Control, 2. ox-LDL alone, 3. ox-LDL+GYY4137 (50 μmol·L⁻¹), 4. ox-LDL+GYY4137 (100 μmol·L⁻¹), 5. ox-LDL+GYY4137 (200 μmol·L⁻¹), 6. ox-LDL+GYY4137 (400 μmol·L⁻¹). (E-b) Bands quantified by densitometric analysis. (F-a) 1. Control, 2. ox-LDL alone, 3. ox-LDL+GYY4137 (200 μmol·L⁻¹), 4. ox-LDL+ZYJ1122 (200 μmol·L⁻¹). Bands quantified by densitometric analysis (F-b). Quantification of ICAM-1 and VCAM-1 mRNA by real-time PCR (G-H). Western blot analysis and quantification of ICAM-1 and VCAM-1 protein expression (I-J). ⁺P < 0.05, compared with no treatment, *P < 0.05, compared with treatment with ox-LDL (n = 3–5).