Figure 2.

Trophoblast organoid-based screening of OPFRs at $\mathrm{10,000}\text{\hspace{0.17em}nM}$. (A) 2- and 4-d culture of trophoblast organoids in screening protocol and scale bars, $100\mathrm{\mu }\mathrm{m}$; (B) Immunofluorescence of villus markers KRT7 (green), GATA3 (red), and DAPI (blue) in trophoblast organoids and scale bars, $30\mathrm{\mu }\mathrm{m}$; (C) Relative fluorescence intensity ($\text{means}\pm \text{SDs}$) of Sytox Green (green) in trophoblast organoids exposed to OPFRs ($\mathrm{10,000}\text{\hspace{0.17em}nM}$) (D) Relative fluorescence intensity ($\text{means}\pm \text{SDs}$) of Ki67 (red) in trophoblast organoids exposed to OPFRs ($\mathrm{10,000}\text{\hspace{0.17em}nM}$); (E) Structure of identified chemicals, EHDPP, o-TCrP, and B4tBPPP. Data in (C) and (D) are expressed relative to the levels in DMSO-treated organoids, which were set to 1. $n=3$. All organoids in (C) and (D) were from a single donor. Data were analyzed using an unpaired two-tailed Student’s $t$-test. Indicated values are significantly different from the control value. Numeric data in (C) and (D) were listed in Table S8. Note: B4tBPPP, bis (4-tertbutylphenyl) phosphate; DAPI, 4′,6-diamidino-2-phenylindole; DMSO, dimethylsulfoxide; EHDPP, 2-ethylhexyl-diphenyl phosphate; GATA3, GATA-binding protein 3; KRT7, keratin 7; OPFRs, organophosphate flame retardants; o-TCrP, tri-o-cresyl phosphate; SD, standard deviation. **$p<0.01$.