Additional file 1 of Hyperactivated Wnt-β-catenin signaling in the absence of sFRP1 and sFRP5 disrupts trophoblast differentiation through repression of Ascl2
Creators: name: Bao, Haili
name: Liu, Dong
name: Xu, Yingchun
name: Sun, Yang
name: Mu, Change
name: Yu, Yongqin
name: Wang, Chunping
name: Han, Qian
name: Liu, Sanmei
name: Cai, Han
name: Liu, Fan
name: Kong, Shuangbo
name: Deng, Wenbo
name: Cao, Bin
name: Wang, Haibin
name: Wang, Qiang
name: Lu, Jinhua
Publisher: name: figshare
Year: 2020
Subjects: subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Neuroscience
subject: Physiology
subject: Biotechnology
subject: Developmental Biology
subject: Cancer
subject: Hematology

description: Additional file 1: : Figure S1. The expression of Sfrp1, Sfrp2 and Sfrp5 during early placental development. A The expression of Sfrp1, Sfrp2, and Sfrp5 was analyzed by whole-mount in situ hybridization at E7.5 and E8.5. B The expression of Sfrp1 and Sfrp5 was detected by in situ hybridization at E7.5-E9.5. The signals were pink. Images in (A) and (B) are representatives of at least two independent experiments. Figure S2. Statistical description of the phenotype of the Sfrp1 and Sfrp5 dKO placenta. A Statistical analysis of spongiotrophoblast thickness and TGCs number in WT and dKO placenta on E9.5. B Quantification of active-β-catenin signal intensity in Fig. 2a. *, P &lt; 0.05. Figure S3. Nuclear localization of active-β-catenin increased in the placenta of Ctnnb1Δ/+ conceptus. A Immunohistochemistry of active-β-catenin in Ctnnb1f(EX3)/+ and Ctnnb1Δ/+ placenta on E9.5. B Quantification of active-β-catenin signal intensity in (A). *, P &lt; 0.05. Figure S4. Global stabilization of β-catenin leads to impaired trophoblast development and embryonic lethality. HE and CK staining of the sections of Ctnnb1f(EX3)/+ and Ctnnb1Δ/+ conceptus on E7.5. Images are representatives of at least three independent experiments. Dec, decidua; EPC, ectoplacental cone; Em, embryo; TGC, trophoblast giant cell. Figure S5. Trophoblast-specific stabilization of β-catenin induces hyperactivation of canonical Wnt pathway. A The localization of active-β-catenin was revealed by immunostaining at E7.5. Cy3-labeled active β-catenin in red, DAPI-labeled nuclei in blue. Images are representatives of at least three independent experiments. B Quantification of active-β-catenin signal intensity in (A). *, P &lt; 0.05. Figure S6. Decreased genes with trophoblast-specific stabilization of β-catenin are related to cell cycles. KEGG analysis of the decreased genes (A, B) between Cyp 19-cre and Cyp19-cre; Ctnnb1f(Ex3)/+ placentas (Fold change&gt;1.5, P value &lt; 0.05). Figure S7. Sfrp1 and Sfrp5 deficiency led to increased activity of Wnt signaling. A TOP-Flash assay in the WT and dKO TS cells. B The expression of Ascl2, Tpbpa and Pl1 in dKO TS cells differentiated for 4 days, in the presence of XAV-939 or not. *, P &lt; 0.05. C QRT-PCR analysis of the expression of Ascl2, Tpbpa and Pl1 in dKO TS cells differentiated for 4 days, with Ascl2 overexpression or not. Figure S8. TCF/LEF might be not required for β-catenin-mediated Ascl2 suppression. A The expression of Ascl2 in WT TS cells treated with indicated conditions. iCRT3, a small molecule that abrogates β-catenin–TCF interaction. *, P &lt; 0.05. ns, not significant. B The top 13 conserved transcription factors binding motif at the β-catenin binding site, 20 kb upstream of the Ascl2 gene. Table S1. Primers Information.