Anti sike

After each treatment, the cells were washed with PBS, fixed in 4% paraformaldehyde for 30 min at room temperature, blocked with 5% (w/v) bovine serum albumin (BSA, Sigma Aldrich, USA) in PBST, and immunostained with anti-Nrf2 (1:100 dilution; Abcam, USA), anti-SIKE (1:100 dilution; Thermo Fisher Scientific, USA), anti-NF-κB (1:100 dilution; Abcam) and anti-p-TBK1 (1:100 dilution; Cell Signaling Technology, USA) antibody overnight at 4 °C, followed by incubation with a goat anti-mouse Alexa Fluor-488-conjugated secondary antibody and/or goat anti-rabbit Alexa Fluor-594-conjugated secondary antibody (Abcam). After washing with PBS, the cells were stained with DAPI (Beyotime, Shanghai, China) and observed under a fluorescence microscope. Immunofluorescence staining for liver sections was performed using anti-Nrf2 (1:100 dilution; Abcam) and anti-SIKE (1:100 dilution; Thermo Fisher Scientific) as described for cells with little modification. Immunofluorescence images were obtained using a fluorescence microscope. Images were analyzed with Image J software.