Th primary antibody

The following reagents were used: In situ Cell Apoptosis Kit produced by F. Hoffmann-La Roche AG; TH Primary Antibody by Santa Cruz Biotechnology in the United States; Bax, Bcl-2, PS, DAB Kit Primary Antibody by Beijing Zhong Shan Golden Bridge Biological Technology CO., LTD; Protease K by Sigma-Aldrich in the United States; SP Kit by Beijing Zhong Shan Golden Bridge Biological Technology Co., LTD. There was no any conflict of interest for all reagents.

The mid brain region of 4% PFA-perfused and sucrose-processed brains were used for immunofluorescence. Tissue sections were taken using Cryotome (Leica CM 1950) and mounted on gelatin-coated slides. The slides containing sections were first incubated in 100% methanol/acetone (chilled at –20°C) at room temperature for 10 min followed by washing with ice-cold PBS (5 min, 3 times), blocking with 3% BSA for a time period of 45 min. The sections were incubated in TH primary antibody (Santa Cruz, 1 : 200 dilution using 1% BSA in PBST) overnight at 4°C followed incubation in Alexa Fluor 488 secondary antibody (Invitrogen, 1 : 2000 dilution using 1% BSA in PBST) for 2 h in the dark at room temperature. The sections were counterstained with fluoroshield DAPI and visualized under the fluorescent microscope (Olympus, BX53) [39 (link)].

The main reagents used in the current study included the following: in situ cell apoptosis kits (Roche, USA), MT (Sigma, USA), TH primary antibody (Santa Cruz Biotechnology, USA), and Bax, Bcl-2, SP, DAB kits, and primary antibody (Beijing Zhongshan Reagents, China). We also used MDA kits that were provided by Boster Biological Technology (Wuhan, China). The following equipment were also used: Synergy-HT multi-function microplate reader (Bio-Tek, USA), ultrasonic homogenizer (Omni, USA), and rat brain stereotactic instrument (NeuroStar, Germany).