Alexa fluor 488 goat anti rabbit igg

Immunofluorescence was performed as previously described in our laboratory25 (link)26 (link). In brief, the tissue sections were deparaffinized, rehydrated and underwent antigen recovery. Then, the sections were permeabilized with 0.4% Triton X-100 for 10 min and blocked using normal goat serum (Zhongshan Golden Bridge Inc., Beijing, China) for 1 h to eliminate nonspecific staining and incubated in a mixture of rabbit anti-NR4A1 antibody (Proteintech), mouse anti-microtubule-associated protein 2 (MAP2) antibody (Zhongshan Golden Bridge) and chicken anti-astrocyte marker glial fibrillary acidic protein (GFAP, Zhongshan Golden Bridge) antibody or goat anti-Aldehyde Dehydrogenase 1 Family Member L1 (Aldh1L1) antibody (Santa Cruz) overnight at 4 °C. Cells were washed using PBS and incubated with Alexa Fluor-350 goat anti-mouse IgG, Alexa Fluor-488 goat anti-rabbit IgG, and Alexa Fluor-594 goat anti-chicken IgG or Alexa Fluor-594 donkey anti-goat IgG (Zhongshan Golden Bridge) in the dark for 2 h at 37 °C. Cells were washed again in PBS, mounted, sealed, and dried overnight. Finally, the images were captured using confocal laser scanning microscopy (Leica, Wetzlar, Germany).

Recombinant Mouse M-CSF and IFN-γ were purchased from R&D. Lipopolysaccharide (LPS) and anti-LC3 antibody were purchased from Sigma Aldrich (St Louis, USA). 2’,7’-dichlorofluorescin diacetate (DCFDA) was from Life Technologies (Carlsbad, USA). Antibodies against NLRP3 were purchased from Cell Signaling Technology. Anti-CD11b (M1/70) and anti-F4/80 (BM8.1) were from BD-Bioscience. Alexa Fluor 488 goat anti-rabbit IgG, anti-mouse IgG and Hoechst 33342 were from Zhongshan Golden Bridge Biotechnology. Anti-NOX2 (ab129068) was purchased from Abcam.