Skip to main content
Fig. 6 | BMC Biology

Fig. 6

From: Hyperactivated Wnt-β-catenin signaling in the absence of sFRP1 and sFRP5 disrupts trophoblast differentiation through repression of Ascl2

Fig. 6

Excessive canonical Wnt signaling activity restrains spongiotrophoblast differentiation through suppressing Ascl2 in cultured trophoblast cells. a The expression of Ascl2 and Tpbpα was analyzed by qRT-PCR in WT as well as Sfrp1 and Sfrp5 dKO trophoblast differentiated for D0-D6. N = 3. *P < 0.05. b Western blot analysis of active-β-catenin during WT and dKO trophoblast differentiation. c Immunostaining of β-catenin (green) and ASCL2 (red) in WT and dKO trophoblast cells at D4. Arrowheads indicated nucleus-located β-catenin. DAPI-labeled nuclei in blue. d, e Immunostaining of β-catenin (green) and ASCL2 (red), and western blot analysis of active-β-catenin in the presence or absence of CHIR at D4. Arrowheads indicated nucleus-located β-catenin. DAPI-labeled nuclei in blue. f The expression of Ascl2 and Tpbpα was detected by qRT-PCR in the presence of CHIR or not, at D4. N = 3. *P < 0.05. gi After treatment with CHIR for 36 h, ChIP-seq showing β-catenin enrichment at Wnt targeted gene loci (Axin2 and Gcm1) (g), and the locus about 20 kb upstream of Ascl2 promoter (h), which is confirmed by ChIP-qPCR analysis (i). N = 3. *P < 0.05. β-actin served as the internal control in b, e. Values are normalized by Gapdh expression level and indicated as means ± SD in a, f, i. Images in c, d are representatives of at least three independent experiments. Scale bar, 100 μm

Back to article page
\