DQ-BSA assay

Approximately 60,000 WT, Irgm1^−/−, Irgm3^−/−, and Irgm1/Irgm3^−/− MEFs were seeded in μ-Slide I chambers. After 24 hours, cells were induced with 200 U/mL IFN-γ or left untreated. After a further 24 hours, cells were incubated with 10 μg/mL DQ Red BSA (Life technologies) [52] and in parallel treated with 200 U/mL IFN-γ or 200 nM Bafilomycin A1 (Sigma-Aldrich) or left untreated. After 1 hour, samples were twice washed with PBS and further incubated in medium containing the corresponding concentration of IFN-γ and Bafilomycin A1. After 3 hours, pictures of live cells on blinded slides were taken and the number of DQ-positive punctae was manually quantified.