Skip to main content
. 2022 Nov 11;13:1035532. doi: 10.3389/fimmu.2022.1035532

Figure 7.

Figure 7

IFNAR2 knockout in wild-type iPS-macrophages recapitulates heightened ZIKV replication and cytopathicity. (A) CRISPR/Cas9 guide design. (B) PCR of IFNAR2 amplicon demonstrating exon 5 splice acceptor site excision in iPSC clones (G8 and G2). Representative of experiments in clones F8 and B5_2. (C) Immunoblot of IFNAR2 and GAPDH expression, demonstrating IFNAR2 ablation in iPS-Mϕ clones (G8, F8, G2, B5_2). Representative of n=2 independent experiments. (D) Expression of macrophage surface markers in IFNAR2 -/- (B5_2) and IFNAR2 +/+ (F8) iPS-Mϕ by flow cytometry, representative of repeat experiments in F8/B5_2. (E) Phagocytic uptake of Zymosan pHrodo particles in IFNAR2 -/- (B5_2) and IFNAR2 +/+ (F8) iPS-Mϕ by flow cytometry, representative of repeat experiments in F8/B5_2. (F) Immunoblot of IFN-I signalling in IFNα2b (1000 IU/mL) treated IFNAR2 -/- (B5_2) and IFNAR2 +/+ (F8) iPS-Mϕ, representative of n = 3 independent experiments. (G) Immunoblot of ENV, RSAD2 and GAPDH expression in IFNAR2 -/- (B5_2) and IFNAR2 +/+ (F8) iPS-Mϕ, 72 h.p.i. post infection, representative of n=3 independent experiments. (H) CellProfiler quantification of cell viability assay in IFNAR2 -/- (G2, B5_2) and isogenic IFNAR2 +/+ (G8, F8) iPS-Mϕ (72 h.p.i. ZIKVFP or ZIKVMP MOI = 1.0, n = 3 independent experiments). Mean ± SD, ANOVA with Sidak’s test for multiple comparisons.