Fig. 3 | Nature Communications

Fig. 3

From: Pre- and peri-implantation Zika virus infection impairs fetal development by targeting trophectoderm cells

Fig. 3The alternative text for this image may have been generated using AI.

Distinct outcomes of dose-dependent ZIKV infection on hESC-derived TECs. a Scheme of ZIKV infection on TECs. b, c Immunostaining of ZIKV E, ZIKV NS2B, KRT7, and hCG in hESCs-derived TECs at 72 h post-infection of MR766 strain ZIKV (MOI = 0.25, Scale bar = 100 µm). Percentage of ZIKV E+ cells was quantified in c. n = 3. d qRT-PCR quantification of ZIKV vRNA in supernatant at 72 hpi of MR766 strain. n = 3. e, f Dose curve of ZIKV infection on day 4 TECs with ZIKV (MR 766, MOI ranging from 10−7 to 101). Intracellular level of ZIKV vRNA at 72 hpi was quantified with qRT-PCR (e); while infectious virus in the supernatant 72 hpi was quantified using Vero assay (f), respectively. n = 3. g–j Immunostaining of ZIKV E and CAS3 in TECs at 72 hpi with high (MOI = 1) and low dose (MOI = 0.001) of MR766 strain ZIKV, Scale bar = 100 µm. The percentage of ZIKV E + cells was quantified in h. Percentage of apoptotic cells was quantified in i. Cell number was quantified in j. n = 3. k Immunostaining for CAS3 of TECs at 72 hpi with ZIKV (MR 766, MOI ranging from 10−7 to 101). l qRT-PCR quantification of intracellular ZIKV ( + ) vRNA in TECs at 72 hpi with high (MOI = 1) and low dose (MOI = 0.001) of MR766 strain ZIKV, n = 3. m Vero assay to quantify ZIKV infectious particles in the supernatant at 72 hpi with high and low dose of MR766 strain ZIKV. n = 3. n Gene Ontology analysis of genes upregulated with ZIKV-infected vs. mock-infected TECs. o qRT-PCR analysis of TECs (left) and trophoblast (right) markers in ZIKV-infected vs. mock-infected TECs as described in g. n = 3. p qRT-PCR analysis of trophoblast marker, CGB, of TECs at 120 hpi with ZIKV (MR 766, MOI ranging from 10E-8 to 10). q Scheme of hNPCs infected with the supernatant harvested at120 hpi with low dose of MR766 strain ZIKV (MOI = 0.001). r–t Immunostaining of ZIKV E in NPCs infected with supernatant of TECs (1:2 dilution) schemed in q. Scale bar = 100 µm. Percentage of ZIKV E + cells was quantified in s and cell number was quantified in t. n = 3. Data are presented as mean ± standard deviation. P values were calculated by unpaired two-tailed Student’s t-test. *P < 0.05, **P < 0.01, and ***P < 0.001, n.s. – not significant. Related to Supplementary Fig. 1 and 2. Source data for 3c–f, h–m, o–p, s–t, are provided as a Source Data file

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