Goat anti rabbit igg h l highly cross adsorbed secondary antibody

The antibodies used in this study included mouse monoclonal anti-CD19 (Cat. AF0093, Beyotime, China), mouse monoclonal biotinylated anti-CD19 (Cat. 302204, Biolegend, USA), rabbit monoclonal recombinant anti-BCMA (Cat. ab253242, Abcam, USA), donkey anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody (Cat. A32766, Invitrogen, USA), goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody (Cat. A-11036, Invitrogen, USA) and Alexa Fluor^® 700 mouse anti-Human CD3 (Cat. 557943, BD Pharmingen™, USA).

Rats were anesthetized with overdosed urethane (2 g/kg i.p., Sigma-Aldrich) and then transcardially perfused with 4% paraformaldehyde in PBS. The brain was removed and immersed in 4% paraformaldehyde for 8 h, and dehydrated with 20% sucrose at 4 °C for 2 days. Coronal sections were sliced into the thickness of 30 μm at MC, STN, SNc and striatum by a frozen microtome. Sections at MC and STN were stained with crystal violet (Nissl stain) to confirm the location of the recording electrode. The sections containing SNc and striatum were washed with PBS, followed by a suppression procedure in 5% fetal bovine serum in 0.1% Triton, and incubated with anti-tyrosine hydroxylase antibody (dilution 1:500, Sigma-Aldrich) overnight at 4 °C, followed by secondary fluorescent antibody (goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary antibody, dilution 1:200, Invitrogen). Images were taken using fluorescence microscope (Zeiss Axiomager, M1). Data were collected only from the animals with confirmed electrode location and dopamine depletion by histological examinations (Supplementary Fig. 4).

HeLa cells grown on polystyrene coverslips (Nest Biotechnology, China) were transfected with the various expression plasmids using PolyJet in vitro DNA transfection reagent (SignaGen, USA) according to the manufacturer's protocol. Cells were washed with PBS at 48 h posttransfection (hpt), followed by fixation in 4% (vol/vol) cold formaldehyde (Beyotime, China) for 15 min at room temperature. The cells were washed with PBS three times after fixation and then permeabilized with 0.1% (vol/vol) Triton X-100 for 15 min, followed by washing with PBS. The cells were blocked with 3% (wt/vol) BSA in PBS for 2 h and then labeled with a mouse (or rabbit) anti-HA antibody (Sigma, USA). The samples were then incubated with a goat anti-mouse IgG (whole molecule) fluorescein isothiocyanate (FITC)-conjugated antibody (Sigma) and a goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody conjugated to Alexa Fluor 633 (Thermo Scientific, USA) in PBS containing 3% BSA for 1 h at room temperature. Nuclei were stained with 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) (Beyotime, China) for 15 min. Images were captured using a Leica DM-IRE2 confocal microscope (Leica, Germany). All experiments were performed in triplicate.