Extended Data Fig. 2: Characterization of Ifnar1−/−/FcγR KO and Ifnar1−/−/ FcγR humanized mice.
From: Human FcγRIIIa activation on splenic macrophages drives dengue pathogenesis in mice
(a) Overview of Ifnar1 gene targeting with CRISPR/Cas9 in mice. Single guide RNA (sgRNA) was designed to target a sequence in exon 3 of the mouse Ifnar1 gene (marked in red). CRISPR-Cas9 constructs were injected to FcγR KO mouse or FcγR humanized mouse and resulted in 1 bp insertion or 1 bp deletion, respectively, that led to a premature stop codon. (b) IFNα/βR expression in various leucocyte populations (see gating strategy on the left) was assessed by flow cytometry in Ifnar1−/−/FcγR KO (grey) or Ifnar1−/−/FcγR humanized (blue) mice in comparison to FcγR humanized mice (red). (c) FcγR expression in various leucocyte populations in the blood was assessed by flow cytometry for Ifnar1−/−/FcγR KO mice (grey) and Ifnar1−/−/FcγR humanized mice (blue) and compared to FcγR expression in FcγR humanized mice. Matching isotype controls are indicated by open histograms. (d) Ifnar1−/−/FcγR KO mice were pre-treated with anti-DENV antibodies and infected with DENV. Pathological changes related to DENV infection were assessed on day 4 post-infection by H&E staining and histological evaluation of mesenteric lymph node (upper panel, 40x), lung (middle panel, 20x) and liver (lower panel, 20x). Images are representative of two infected mice.
