Siparkin

SiRNAs against PINK1 (siPINK1) (5′‐GCUAGUUACAAGAGAACAA‐3′), Parkin (siParkin) (5′‐GAAUACAUUCCCUACCUCA‐3′), Nrf2 (siNrf2) (5′‐GAGAAAGAAUUGCCUGUAA‐3′), and HO‐1 (siHO‐1) (5′‐CAGUUGCUGUAGGGCUUUA‐3′) and scrambled siRNA (si‐Control) (5′‐UUCUCCGAACGUGUCACGU‐3′) were designed and synthesized by RiboBio. Nucleus pulposus cells were transfected for 48 hours with 100 nmol/L of each siRNA using Lipofectamine® 2000 (Invitrogen) according to the manufacturer's instructions. The cells were then used for experiments.

A lentiviral vector containing rat Park7 (OE-Park7) and a lentiviral vector containing rat Hk2 (OE-Hk2) were constructed by Obio Technology (China). shRNAs targeting rat Park7 (shPark7) and Hk2 (shHk2) and a negative control shRNA (shControl) were synthesized and inserted into lentiviral vectors by Obio Technology. Transfection was performed according to the manufacturer’s instructions. For siRNA transfection, the siRNA targeting rat Prkn (si-Parkin) and the control siRNA (si-Control) were synthesized by RiboBio (China). siRNA transfection was performed using riboFECT™ CP transfection reagent (RiboBio, China) according to the manufacturer’s instructions.