Fig. 1

TGFBIp transcription and H3K4me3 levels in wild-type and GCD2-homozygous corneal fibroblasts. a, b Slit-lamp photographs and pedigree of wild-type, GCD2 heterozygous, and GCD2 homozygous cells. c, d The mRNA and protein levels of TGFBIp in normal and GCD2 corneal fibroblasts was determined by RT-qPCR (c) and western blot (d). Gene expression was normalized to internal control GAPDH gene, and results are expressed as fold stimulation over control. e, f H3K4me3 and H3K27me3 levels at the Smad binding elements on TGFBIp gene promoters and at the TSS of TGFBIp gene in wild-type and GCD2-homozygous corneal fibroblasts. ChIP assays were performed with H3K4me3 and H3K27me3 antibodies. Immunoprecipitated DNA and input DNA were subjected to qPCR with primers specific for the TGFBIp gene promoter to measure enrichment levels. qPCR data were analyzed using the 2^-ΔΔCt method, and data are the mean fold change relative to the input ± standard error (SE) of enrichment. (mean ± SE; **P < 0.01 vs. wild type, n = 3)