Analysis of the role of ZEB1 in the pathogenesis of posterior polymorphous corneal dystrophy.

ABSTRACT:

Purpose

To determine how nonsense mutations in the transcription factor ZEB1 lead to the development of posterior polymorphous corneal dystrophy type 3 (PPCD3).

Methods

Whole-cell extracts were obtained from cultured human corneal epithelial cells (HCEpCs) as a source of ZEB1 protein. DNA-binding assays were performed using the whole-cell extract and oligonucleotide probes consisting of the two conserved E2-box motifs and surrounding nucleotides upstream of COL4A3. ZEB1 and COL4A3 mRNA expression in primary human corneal endothelial cells (HCEnCs) was assayed in both PPCD3 and control corneas by RT-PCR. Immunohistochemistry was used to localize ZEB1 and COL4A3 expression in normal human cornea.

Results

Electromobility shift assays (EMSAs) and competition EMSAs demonst

SUBMITTER: Yellore VS 

PROVIDER: S-EPMC3292363 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature