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. 2022 Nov 11;13:1035532. doi: 10.3389/fimmu.2022.1035532

Figure 9.

Figure 9

Enhanced ZIKV infection and CPE in IFNAR2-deficient iPS-microglia-like cells. (A) Immunofluorescence analysis of microglial markers IBA1 and TMEM119 in IFNAR2 PT (PT clone 11) and IFNAR2 WT (WT2) iPS-Mϕ and iPS-microglia-like cells (iPS-MGLs), representative of repeat experiments in G2, B5_2, G8 and F8 lines. Scale bar = 100 μm. (B) Immunoblot of ENV, RSAD2, ISG15, GAPDH in ZIKVFP-infected IFNAR2 +/+ (F8) and IFNAR2 -/- (B5_2) iPS-MGLs at 72 h.p.i., representative of n = 3 independent experiments in IFNAR2 PT [clones 6 and 11] and IFNAR2 WT [WT2]). (C) Plaque assay on Vero cells of ZIKV infectious particles in supernatants (48 h.p.i. ZIKVFP MOI = 1, n = 3 independent experiments in IFNAR2 WT (WT2) and IFNAR2 PT (clone 6) iPS-microglia-like cells. Mean ± SD, t test. (D) RT-PCR quantification of IL6, IL1B and TNF relative to 18S (24 h.p.i. ZIKVFP MOI = 1, n = 3 independent experiments in IFNAR2 WT [WT2] and IFNAR2 PT [clones 6 and 11]) microglia-like cells. Mean ± SD, ANOVA with Sidak’s test for multiple comparisons. (E) Cell viability assay (72 h.p.i. ZIKVFP MOI = 1.0 [n = 4 independent experiments] or ZIKVMP MOI = 1.0 [n = 3 independent experiments]) in IFNAR2 -/- (B5_2), IFNAR2 PT (clone 6), IFNAR2 +/+ (F8) and IFNAR2 PT (WT2) iPS-MGLs. Mean ± SD, ANOVA with Sidak’s test for multiple comparisons.