Nuclear protein extraction kit

Total proteins were extracted from the rat pancreas, while pancreatic nucleoproteins were extracted by using a Nuclear Protein Extraction Kit (Jiancheng, Nanjing, China). Protein concentrations were quantified using a BCA protein assay kit (Pierce BCA). Equal amount of proteins were separated on 8% SDS-PAGE gels and transferred onto the nitrocellulose membranes. The membranes were then incubated overnight at 4 °C with the following primary antibodies: Nrf2(dilution 1:1000, proteintech), Keap1(dilution 1:1000, CST), inhibitor of nuclear factor kappa B kinase β(IKKβ, dilution 1:1000, CST), NF-κB p65(dilution 1:1000, CST), phosphorylated phosphatidylinositol-4,5-bisphosphate 3-kinase(p-PI3K, dilution 1:1000, CST), phosphorylated protein kinase B(p-AKT, dilution 1:1000, CST), GAPDH (dilution 1:5000, proteintech), and laminB1(dilution 1:2000, Abcam). After incubated with the corresponding secondary antibodies for 2 h, the protein bands were visualized by using the Odyssey scanner (LI-COR Biosciences).

Z. bungeanum Maxim. (batch number 20180802) was purchased from Hehuachi tradition herbal medicine market, Chengdu, China. Mesylate dihydroergotoxine tablets (batch number 7K893T) were purchased from Tianjin Huajin Pharmaceutical Co., Ltd., Tianjin, China. D-galactose (batch number 2017060101) was purchased from Chengdu Cologne Chemical Co., Ltd., Chengdu, China. Superoxide dismutase (SOD) kit, malondialdehyde (MDA) kit, glutathione (GSH) kit, catalase (CAT) kit, and nuclear protein extraction kit were obtained from Nanjing Jiancheng Bioengineering Institute, Nanjing, China (batch number 20180605, 20180611, 20190718, 20190719, and 20190722, respectively). Primary antibodies, including PI3K (Cat no.: 4257S), AKT (Catno.: 4685S), p-PI3K (Cat no.: 4685S), p-AKT (Cat no.: 4060S), and Lamin A/C (Cat no.: 4777T) were provided by Cell Signaling Technology, MA, USA; HO-1 (Cat no.: 10701-1-AP), Bcl2 (Cat no.: 26593-1-AP), and Bax(Cat no.: 50599-2-1g) were provided by Wuhan Sanying Biotechnology Co., Ltd., Wuhan, China. Nrf2 (Cat no.: ab34436-050) was provided by Hangzhou Lianke Biotechnology Co., Ltd., Hangzhou, China.