Fig. 5
From: Male germ cells support long-term propagation of Zika virus
BC inhibits ZIKV infection of murine male GCs in vitro and in vivo. a Hit compounds that inhibit ZIKV infection in GCs by more than 30%. b Chemical structure of BC. c Inhibition curve of BC. IC50 = 2 μM. d Intracellular flow cytometry plots of GCs at 72 hpi with ZIKV (MOI = 0.1 PFU per cell) in the presence of 30 μM BC (n = 6). e–h qRT-PCR analysis of relative ZIKV cellular (e) and negative-strand (f) vRNA in GCs, vRNA in supernatant (g), and infectivity with Vero assay (h) in the presence of 30 μM BC. i, j BC significantly reduced ZIKV infection in long-term infected GC. Culture media containing BC were refreshed every 24 h (starting at 70 dpi) and assessed for relative ZIKV vRNA in the supernatant by qRT-PCR (i) and for infectivity with Vero assay (j). k Schematic representation of BC treatment time course for (l–p). l Relative percentage of ZIKV + GCs in the presence of 30 µM BC (n = 9), measured by intracellular flow cytometry. m–p qRT-PCR analysis of relative ZIKV cellular (m) and negative-strand (n) vRNA in GCs, vRNA in supernatant (o), and infectivity with Vero assay (p). q Infectivity of ZIKV after incubation with 30 µM BC for 24 h, measured by Vero assay. Samples were diluted 1:1,000. r qRT-PCR analysis of ZIKV vRNA in testes of BC-treated and vehicle-treated ZIKV-infected mice at 6 dpi (n = 18 for mock-infected, 18 for ZIKV-infected vehicle-treated, and 20 for ZIKV-infected BC treated). s qRT-PCR analysis of ZIKV negative-strand vRNA in testis of intratesticular mock-infected and ZIKV-infected mice at 1 dpi (n = 10 per group). t, u qRT-PCR analysis of ZIKV cellular (t) and negative-strand (u) vRNA in intratesticular ZIKV-infected mouse testis after BC or saline treatment 1 to 4 dpi (n = 10 per group). e–j, l–q Data are normalized to values obtained from DMSO-treated groups. n = 3 for all in vitro experiments, unless otherwise indicated. Statistical values are presented as mean ± s.e.m. and analyzed by two-sided t test and one-way ANOVA, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, or no significance (ns). ZIKV strain is MR 766 throughout
