Fig. 1: Wnt/β-catenin signaling activates Nlgn3 transcriptional program in hippocampal cells.

a Top: Genomic context of human NLGN3 in the long arm of chromosome X and schematic exon-intron boundaries of the gene. White and gray boxes: 5′ and 3′ UTR and exons, respectively. Middle: Conservation profile of the human NLGN3 promoter sequence compared with similar genomic regions in Mus musculus and Rattus norvegicus (50–100%). Bottom: Schematic representation of potential TCF/LEF sites (TBE: CTTTG, circles) found in these species. ECR: Evolutionary conserved region. b Early expression levels of Nlgn3 and cMyc genes after 2 h treatment with increasing doses of either purified Wnt3a protein or LiCl in HT22 hippocampal cells or rat primary hippocampal neurons (RHN). Rpl13a was used as a reference gene. c Quantitative determination of Nlgn3 and cMyc mRNA levels after 2 h treatment with Wnt3a (200–400 ng/ml) protein or LiCl (10–20 mM) in HT22 hippocampal cells and RHNs. d TOP: protein levels of β-catenin after 48 h treatment with β-catenin-shRNA. shRNA of GFP was used as control and β-actin as a loading control. Bottom: expression levels of Nlgn3 and β- catenin. e Nlgn3 and β-catenin protein levels in HT22 cells and RHNs after 6 h treatment with 200 and 400 ng/ml of purified Wnt3a protein. β-actin was used as a loading control. In (c) and (d), data represent mean ± s.e.m., *P < 0.05, **P < 0.01, two-tailed Mann–Whitney test