Rabbit anti nrf2 polyclonal antibody

Brain tissues and astrocytes were treated according to experimental designs. WB was performed as previously described (Song et al., 2019 (link)). The following primary antibodies were used: rabbit anti-Nrf2 polyclonal antibody (1:500, Santa Cruz Biotechnology), rabbit anti-HO-1 polyclonal antibody (1:2000, Abcam), mouse anti-Connexin43 monoclonal antibody (1:1000, Invitrogen), rabbit anti-Cx43 polyclonal antibody (1:300, Sigma-Aldrich), rabbit anti-PKCα polyclonal antibody (1:1000, ABclonal Technology, China), rabbit anti-phospho-PKCα polyclonal antibody (1:1000, ABclonal Technology), rabbit anti-GFAP polyclonal antibody (1:1000, Servicebio), mouse anti-VIM monoclonal antibody (1:1000, Servicebio), rabbit anti-histone-H3 monoclonal antibody (1:1000, Cell Signaling Technology, United States), and mouse anti-GAPDH monoclonal antibody (1:2000, Servicebio). Enhanced chemiluminescence solution (Thermo Fisher Scientific) and Tanon Image (Shanghai, China) were used to detect the chemiluminescence signal. The relative intensity of the bands was measured by ImageJ 1.6.0 (NIH, United States).

Cells were plated onto coverslips, handled according to experiment design. The cells were fixed in 4% PFA for 10 min. After washing with PBS three times, fixed cells were permeated with 0.3% Triton X-100 for 10 min, and blocked with 1% bovine serum albumin (BSA) for 1h at room temperature, then incubated overnight at 4°C with primary antibodies: rabbit anti-Nrf2 polyclonal antibody (1:200, Santa Cruz Biotechnology, United States) and rabbit anti-HO-1 polyclonal antibody (1:300, Abcam, United Kingdom). After washing with PBS three times, the cells were incubated with corresponding secondary fluorescent antibodies (1:300, Invitrogen, United States): Alexa Fluor 488 donkey anti-rabbit IgG for 1 h at room temperature. And then the nuclei were counterstained with DAPI (1:5000, Beyotime Biotechnology, China). A confocal laser-scanning microscope (Leica, Germany) was used to observe and analyze fluorescence images.