Fig. 4: ILF3 knockout accelerates ZIKV-associated microcephaly in brain organoids and mice model. | Nature Communications

Fig. 4: ILF3 knockout accelerates ZIKV-associated microcephaly in brain organoids and mice model.

From: Identification of antiviral RNAi regulators, ILF3/DHX9, recruit at ZIKV stem loop B to protect against ZIKV induced microcephaly

Fig. 4

A Demonstrative Brain organoids and size measurement in mock or ZIKV-infected control or ILF3-depleted brain organoids (MOI = 1) at day 15 (scale bar, 200 μm). Upper panel: schematic diagram of brain organoid protocol. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. n.s., non-significance, *p ≤ 0.05, **p < 0.01. B RT-qPCR showing ILF3 mRNA expression and viral RNA copies in mock or ZIKV-infected control or ILF3-depleted brain organoids (MOI = 1) at day 15. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. **p < 0.01, ***p ≤ 0.001. C Flowchart demonstrated the experiment procedure of ZIKV-associated microcephaly control or Nestin-Cre ILF3 cKO mice model. D Survival rate. Lethality was monitored for 10 days. Kaplan–Meier survival curves were analyzed by the log-rank test, **p < 0.01, n = 8 mice/group. Representative image of mock or ZIKV-infected WT or Nestin-Cre ILF3 conditional knockout mice (cKO) mice (2 × 105 PFU/mice), brain size (E), and brain weight (F) captured at 7 dpi. Bars, mean ± SD, n = 8 mice/group, two-tailed Student’s t-test. n.s., non-significance, *p ≤ 0.05, **p < 0.01. G Clinical score measured in mock or ZIKV-infected control or ILF3 cKO mice (2 × 105 PFU/mice) for 7 days. Bars, mean ± SD, n = 8 mice/group, one-way ANOVA analysis. *p ≤ 0.05, **p < 0.01. H Immunofluorescence showed the relative expression of GFP, SOX2, and ZIKV-E in mock or ZIKV-infected control or ILF3 cKO mice (scale bar, 100 μm). Bars, mean ± SD, n = 3 mice, two-tailed Student’s t-test. **p < 0.01. I RT-qPCR showing viral RNA copies in ZIKV-infected control or ILF3-cKO mice brain at 7 dpi. Bars, mean ± SD, n =6 mice/group, two-tailed Student’s t-test, **p < 0.01. J Virus titers were quantified by virus plaque assay from ZIKV-infected WT or ILF3-cKO mice brain at 7 dpi. Bars, mean ± SD, n = 6 mice/group, two-tailed Student’s t-test, **p < 0.01. K Hematoxylin and eosin stain (HE) staining of the hippocampus of mock and ZIKV-infected mice brain (Scale bar, 100 μm). Red arrow: infiltrated mononuclear cells. n = 3 mice. Source data are provided as a Source Data file. C was created with BioRender.com.

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