Fig 2.

Verification of transgene expression in the retina. (A) For the verification of the expression of the transgene in the retina, mRNA samples from retinal extracts were transcribed via reverse transcription and subsequently amplified via PCR. As controls mRNA samples were used as PCR template to check them for contaminations of genomic DNA, which could lead to inaccurate test results. (B) PCR products were checked for the mutational sites using direct Sanger sequencing. The comparison of the histograms shows the pathological point mutation (an adenine deletion sequenced on the complementary strand and therefore displayed as thymine deletion.) (C) Western blot of retinal extracts (∼100 μg) from wild type (wt) and mutant (ty), and as negative control from Rpgr^ko (KO) mice at the age of 3 months probed with the RPGR^ORF15 specific antibody. Immunoblot of the same samples with anti-β-tubulin antibody served as loading control. DNA, deoxyribonucleic acid; PCR, polymerase chain reaction; RPGR, retinitis pigmentosa GTPase regulator.