Fig 1: The shRNA sequence of WNT3 gene
Fig 2: The expression levels of WNT3 gene in placentas of the three group and its correlation with methylation levels. A The mRNA expression levels of WNT3 in placentas in the three groups. B The correlation between mRNA expression and methylation levels. r = - 0.525, R2 = 0.2071, P < 0.05. C, D The relative protein expression of WNT3 in placentas of the three groups by western blotting. TB term birth, PB preterm birth, PE preeclampsia. Unpaired t test, Welch's t test, Mann–Whitney test, and Pearson correlation analysis. *P < 0.05, **P < 0.01. ***P < 0.001, ****P < 0.0001
Fig 3: Loss of WNT3 suppressed the Wnt/β-catenin signaling pathway. A Expression of Wnt/β-catenin signaling pathway proteins in HTR8/SVneo cell line after transfection with shRNA. 1–3, Control; 4–6, sh-WNT3 (cells were transfected with shRNA-WNT3); 7–9, sh-NC. B The relative protein expression of WNT3, β-catenin, phosphorylated β-catenin, GSK3β, phosphorylated GSK3β, and ratio of phosphorylated and non-phosphorylated protein in HTR8/SVneo cell line. One-Way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001
Fig 4: WNT3 promoted trophoblast cell proliferation. A Cell Counting Kit-8 assay was used to detect the proliferation of HTR8/SVneo cells. B Cell proliferation increased after transfection with WNT3-plasmid in HTR8/SVneo cells. One-way ANOVA. *P < 0.05, **P < 0.01.Image acquisition tools: IBM SPSS Statistics 25 and GraphPad Prism 8.4.2. Image processing software packages: Adobe photoshop cs6 13.0.1
Fig 5: The methylation levels of WNT3 gene by pyrosequencing. A The average levels of the four CpGs in PE (12.4% ± 2.2%) compared with TB (14.5% ± 2.2%) and PB (14.3% ± 2.5%) groups. B–E The methylation levels of the four CpGs. TB term birth, PB preterm birth, PE preeclampsia. Unpaired t test. *P < 0.05, **P < 0.01
Supplier Page from Abcam for Anti-Wnt3 antibody