Material and methods
Chemicals and reagents
DIOS (purity 98% by HPLC) is provided by the Chemistry Institute of
Pharmaceutical Resource of Southwest University (China). The mouse
insulin ELISA kit was purchased from Shanghai enzyme-linked
Biotechnology Co., Ltd (ml001827-1). The primary antibodies used for
western blot were: IRS (CST#2383, 1:1000), p-IRS
(Ser307, CST#2381, 1:1000), PI3K
(CST#4257, 1:1000), PI3Kα (CST#4255, 1:1000), AKT (CST#3063, 1:1000),
p-AKT (Ser473, CST#4060, 1:1000), GSK-3β (CST#9315, 1:1000), p-GSK-3β
(Ser9, CST#5558, 1:1000), GS (CST#3886, 1:1000), AS160 (CST#2670,
1:1000),p-AS160 (Thr642, CST#8881, 1:1000), Glut4 (CST#2213, 1:1000),
β-actin (Proteintech 20536-1-AP, 1:5000) and secondary antibody (cat.
#SA00001-2, 1:10000) were purchased from Proteintech (Wuhan, China),
and total protein extraction kits are from Sangon Biotech Co. Ltd
(Shanghai, China).