Figure 1: Screening GWAS genes identifies critical negative regulatory nodes controlling macrophage oxidative burst.

(a) Workflow of primary screen. Genes associated with IBD and autoimmunity were screened to identify regulatory nodes controlling oxidative burst in RAW264.7 macrophages. Gene knockdown was achieved by lentiviral transduction with shRNAs, and cells were stimulated with IFN-γ and zymosan to induce oxidative burst as measured kinetically by luminol chemiluminescence. (b) For each hairpin tested, oxidative burst was quantified as the area under the curve (AUC) derived from kinetic luminescence plots, which was normalized to cell number (Alamar Blue) and ultimately expressed as Z-scores. (c) Among >400 genes screened, hits were classified as either activators or inhibitors of oxidative burst by using Ncf4 and Cat as phenotypic references. FPR, false positive rate. (d) We identified four novel negative regulators (inhibitors) of the oxidative burst with robust false positive rates. See also Supplementary Data 1.