Fig. 5. Ouabain protected pregnant C57BL/6 mice and their developing fetuses.

a Schematic depiction of ouabain treatment. b E13.5 uteri from ouabain-treated and vehicle-treated anti-Ifnar dams. c Representative images of fetuses from ouabain and vehicle-treated group. d Fetus survival on E11.5–13.5 after infection with ZIKV at E6.5–8.5. Data are representative of at least three independent experiments with one pregnant female dam per experiment. The n for each group is indicated above each bar. **P < 0.01. e Fetus size, as assessed by crown–rump length × occipital–frontal diameter at day 5 post-infection. Bars indicate the mean size of 7–10 fetuses from three independent experiments from fetuses carried by 2–3 pregnant dams. ****P < 0.0001. f, g qRT-PCR analysis of viral burden in fetal head and placenta on E11.5–13.5 after infection at E6.5–8.5. Symbols represent individual fetuses collected from several independent experiments. Dashed lines, limit of detection. *P < 0.05; ***P < 0.001. h Ouabain alleviated reductions in the size of the labyrinth layer of placentas caused by ZIKV infection. i Ouabain blocked the loss of trophoblasts caused by ZIKV infection. Immunofluorescence staining of cytokeratin (CK) in mouse placentas. j–l Ouabain protected embryonic brains from ZIKV infection and microcephaly. Low magnification images of hematoxylin and eosin (H/E) staining of vehicle- and ouabain-treated fetuses on E13.5 (j). High magnification images of H/E staining of cerebral cortex of vehicle- and ouabain-treated fetuses on E13.5 (k). Images of cortices stained for NPCs (green, SOX2) and DAPI (blue) (l).