Fig. 6: Impact of maternal Zika infection on offspring NETosis.

a Venn diagram of top pathways unique to female and male offspring bone marrow neutrophils after in utero ZIKV exposure. Analysis was performed using DESeq2 with Benjamini–Hochberg correction for multiple comparisons. All tests were two-sided. b Heatmap of DEGs related to “Neutrophil-mediated extracellular trap formation.” c mRNA expression of MPO and Elane in neutrophils from male and female offspring in control and ZIKV-exposed groups. For males: control (n = 5 isolations) and ZIKV (n = 4 isolations). For females: control (n = 4 isolations) and ZIKV (n = 4 isolations). Each isolation represents neutrophils pooled from the bone marrow of two mice, with each data point in the graph corresponding to one isolation. d Experimental design. Created in BioRender. Alvero, A. (2025) http://BioRender.com/bb2qwff. Bone marrow neutrophils were isolated from male and female offspring on PND35 from control and Zika-exposed groups and were treated with phorbol 12-myristate 13-acetate (PMA) to evaluate NETosis. e Representative image of NET formation. Red: MPO; green: SYTOX, indicating extracellular DNA. Co-localization of SYTOX Green and MPO confirmed the presence of NETs. Scale bar = 130 µm. f ROS production of isolated neutrophils when treated with PMA for 45 min. Neutrophils were isolated from 2–4 mice per experiment across 3 independent experiments. ****p < 0.0001. g Heatmap of DEGs related to “Reactive Oxygen Species Metabolic Process.” Data are shown as the mean ± SD. Two-sided Student’s t-test (c) and Two-way ANOVA with Šídák’s multiple comparisons (f) were used. FC female control neutrophils, FZ female ZIKV neutrophils, MC male control neutrophils, MZ male ZIKV neutrophils, ROS reactive oxygen species. Female and male mice symbols (panel e and f) were created in BioRender. Alvero, A. (2025) http://BioRender.com/u0w40ys. Source data are provided as a Source Data file.