Anti rat fitc conjugated antibody

Pyroptotic BMDMs were fixed for 1 hour at room temperature (RT) in 4% paraformaldehyde (PFA) (Electron Microscopy Sciences, 15710), then incubated with PBS containing 50 mM NH 4 Cl (Sigma, A9434) for 10 min and washed in the same buffer. Samples were incubated in PBS containing 10 µg/mL of anti-mouse CD68 (Biolegend, 137001) or IgG2a isotype control (Biolegend, 400501) and 0.25% of gelatin for 1 hour at RT. After a washing step, samples were incubated in a solution containing 10 µg/mL of anti-Rat FITC conjugated antibody (Jackson ImmunoResearch, 712-096-153) and 0.25% of gelatin for 1 hour at RT. Finally, samples were washed in PBS, incubated in PBS containing 10 µg/mL of Hoechst 33342 (Invitrogen, H3570) for 10 min at RT, and coverslips were mounted in Mowiol following further washing steps in PBS and distilled water. Images were acquired using the UltraView VOX microscope and data were analysed using the FIJI software.