Additional file 1: of The manner of decay of genetically defective EYS gene transcripts in photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients depends on the type of mutation

Table S1. Primer sequences for RT-PCR. Figure S1. Cell growth of dermal fibroblasts with or without EYS gene defects. The number of cells increased until about 14 days; however, growth rates were different depending on the donor. The highest was N#1, followed in order by Pt#2, N#3 and Pt#1 (N#1>Pt#2>N#3≒Pt#1). Although the results suggest that the cell growth may be determined mainly by donor age and defects in the EYS gene are only relevant for younger EYS-RP patients, a larger sample size should be tested to assess differences in cell growth between EYS-RP patients and normal volunteers. Figure S2. Exogenous expression of transgenes, CRX, RAX, NeuroD and OTX2, in induced photoreceptor-like cells derived from fibroblasts of 3 arRP patients and 3 normal volunteers. Exogenous expression of 4 transgenes, CRX, RAX, NeuroD and OTX2 initiated 10 days after gene transduction. Figure S3. Expression of CRX and blue opsin in induced photoreceptor-like cells derived from fibroblasts of 3 arRP patients and 3 normal volunteers. Immunocytochemistry using antibodies to CRX and blue opsin (green). Figure S4. Induction of retina-specific genes, blue opsin and S-antigen, in human dermal fibroblasts by the retroviral infection of genes for defined transcription factors. Figure S5. Expression levels of EYS gene transcripts over time during photoreceptor-directed differentiation by qRT-PCR. a. Changes of expression levels of EYS gene transcripts over time during photoreceptor-directed differentiation by qRT-PCR. Expression levels of the EYS gene in Pt#1, Pt#2 and N#2 reached maximum levels around 10 to 14 days after gene transduction and remained unchanged for up to 3 weeks. b. Statistical analysis of expression levels of EYS gene corresponding to exon 8-9 by qRT-PCR. Expression levels of exon 8-9 of Pt#1 and Pt#2 were almost the same as the average of three normal volunteers 10 days after gene transduction (Wilcoxon test, p = 0.1747 for Pt#1 and p = 0.1725 for Pt#2). Figure S6. The inserted sequence between exon 26 and exon 27. (PDF 846 kb