P iκb β

The protein from RAW264.7 cell samples was directly extracted based on the manufacturer's standard protocols (Beyotime Biotechnology). The cytoplasmic and nuclear protein fractions were extracted using Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime Biotechnology) according to the manufacturer's protocol. Cytoplasmic/nuclear protein extracts or whole protein extracts were used for Western blot analysis. The primary rabbit antibodies for nuclear factor kappa B (NF-κB) p65 subunit (p65), phosphorylated NF-κB p65 (p-p65), phosphorylated inhibitor of kappa B alpha (p-IκBβ), and phosphorylated IκB kinase-β/β (p-IKKβ/β) (Cell Signaling Technology, Boston, USA) proteins were used for the detection of p65, p-p65, p-IκBβ, and p-IKKβ/β protein expression. Moreover, the primary rabbit antibodies for proliferating cell nuclear antigen (PCNA), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), IKKβ, IKKβ, IκBβ proteins (Cell Signaling Technology) were used for the detection of PCNA, GAPDH, IKKβ, IKKβ, and IκBβ proteins, which were used as the control. Immunostained proteins were detected by electrochemiluminescence (CWBio Co., Ltd., Beijing, China).