Fig. 7: ZIKV^E67A mutant virus exhibits significantly lower inhibition on steroidogenesis and spermatogenesis than ZIKV^WT.

a Viral RNA copy numbers in ZIKV-infected mouse testes (days 6) were quantified using qRT-PCR. b–e Testes were infected with ZIKV^WT or ZIKV^E67A for 6 days. The levels of testosterone and progesterone in the serum (b, c) or testis homogenates (d, e) were tested by ELISA. f, g Protein levels of CYP17A1/CYP11A1/STAR/3β-HSD and ZIKV-NS2A in ZIKV-infected mouse testes (f), and CYP17A1 mRNA levels (g) were measured by Western blotting and qRT-PCR, respectively. h Representative immunofluorescence images of ZIKV-infected mouse testes by staining for CYP17A1 and NS2A. DAPI staining indicates the nuclei. Scale bar, 100 μm. i, j Computer-assisted sperm analysis [total (i) and motile (j)] on samples obtained from the cauda epididymis of ZIKV-infected mice immediately after euthanasia. k Representative immunofluorescence images of ZIKV-infected mouse testes by staining for DDX4. DAPI staining indicates the nuclei. Scale bar, 100 μm. l Histological analysis of the epididymis collected from ZIKV-infected mice stained with hematoxylin and eosin. Stars indicate loss of spermatogenic cells in testis. Scale bars, 100 μm. The images in f, h, k, l are representative of three independent experiments. The ocular lens was set to 15× in panel h, and to 10× in panels k and l, with the magnification values corresponding to the objective lens used. The data shown in a–e, g, i, j are the mean ± s.d (n = 5 independent experiments); Statistical analysis of (a–e, g, i, j) was performed using one-way ANOVA, followed by Tukey’s multiple comparison test; ns, not significant. Source data are provided in the Source Data file.