Supplementary Figure 3: Expression analysis of the ABCA1, AFAP1 and GMDS genes in human ocular tissues.
From: Common variants near ABCA1, AFAP1 and GMDS confer risk of primary open-angle glaucoma
(a-d) Expression of the ABCA1 (a), AFAP1 (b,c) and GMDS (d) mRNA was analyzed by RT-PCR using gene specific primers. (a) The 913 bp PCR product amplified with ABCA1-specific primers in iris, ciliary body (CB), retina, optic nerve head (ONH), optic nerve (ON) and normal (NTM) and glaucomatous (GTM) trabecular meshwork cell lines (respectively, NTM-5 and GTM-3) corresponds to the expected size of the predominant transcript encoded by the gene. The 770 bp amplicon in the ciliary body, retina, optic nerve head and normal and glaucomatous trabecular meshwork cells represents the alternate transcript lacking exon 4. The origin of the >1000 bp amplified product in the ciliary body and retina is unclear. (b) The 408 bp PCR product amplified with AFAP1-specific primers in the indicated tissues corresponds to the expected size of the ubiquitously expressed B isoform. (c) Upon increasing the amplification cycles, in addition to the 408 bp amplicon, the 660 bp product in the retina corresponds to the neuronal cell specific, A isoform of AFAP1. The product marked with an asterisk was found to be non-specific by sequencing. Amplification from three independent cDNA preparations is shown. (d) The ∼300 bp PCR product amplified with primers specific to the GMDS variant 1, in all the analyzed tissues corresponds to the expected size of this variant. Representative data from two independent experiments are shown. M, molecular size markers; RT-, without reverse transcriptase control; bp, base pair.
