Tissue cryosections were fixed in 4% PFA at 4°C
overnight, embedded in optimal cutting temperature (OCT) compound, and sectioned
in a cryostat (12 μm). For IL-1β immunostaining, sections were
incubated with a polyclonal goat anti-mouse IL-1β primary antibody
(1:200, R&D Systems) and monoclonal rat anti-mouse CD11b antibody (1:500,
AbD Serotec). After washing 3 times with TBST, sections were incubated with
Cy3-labeled donkey-anti-rat IgG antibody (1:200, Jackson Lab) and Alexa
488-labeled donkey anti-goat IgG antibody (1:200,Molecular Probes). Sections
were mounted with mounting medium containing DAPI (Invitrogen), and confocal
microscopy was performed.
overnight, embedded in optimal cutting temperature (OCT) compound, and sectioned
in a cryostat (12 μm). For IL-1β immunostaining, sections were
incubated with a polyclonal goat anti-mouse IL-1β primary antibody
(1:200, R&D Systems) and monoclonal rat anti-mouse CD11b antibody (1:500,
AbD Serotec). After washing 3 times with TBST, sections were incubated with
Cy3-labeled donkey-anti-rat IgG antibody (1:200, Jackson Lab) and Alexa
488-labeled donkey anti-goat IgG antibody (1:200,Molecular Probes). Sections
were mounted with mounting medium containing DAPI (Invitrogen), and confocal
microscopy was performed.