Figure 3
From: Blepharocheilodontic syndrome is a CDH1 pathway–related disorder due to mutations in CDH1 and CTNND1
Functional analysis of CDH1 mutations. (a) E-cadherin expression levels in cells expressing wild-type (wt) and mutated CDH1. The HDGC-associated mutation p.(Asp244Gly) is used as a positive control. There is no detectable E-cadherin protein for mutations p.(Asp254Tyr) and del exon 9 (c.1320G>T and c.1320 + 1G>C inducing exon 9 skipping). (b) Representation of the relative quantification of total E-cadherin. Compared to wild type, substitution p.(Val454del) shows a dramatic decrease in E-cadherin protein expression (−50%; P = 0.05), whereas substitution p.(Asp257Val) shows only a slight decrease (−6%; P = 0.05). Substitution p.(Asp244Gly) showed equal amount of E-cadherin protein compared to wild type. Significant P-values are indicated by * (P < 0.05). (c) Confocal microscopy images of HEK239T cells expressing wt and mutated CDH1 double-labeled with anti-E-cadherin antibody (green) and p120ctn antibody (red). HEK293T cells do not express detectable E-cadherin. In CDH1 wt-expressing cells, E-cadherin is mainly located at the plasma membrane, colocalizing with p120ctn. Mutations identified in CDH1 induce loss of cytoplasmic membrane staining and p120ctn colocalization and show intracytoplasmic perinuclear accumulation. The HDGC-associated mutation p.(Asp244Gly) also induced loss of cytoplasmic membrane localization and intracytoplasmic E-cadherin accumulation.
