Figure 1

Transgenic mice expressing GARP2 in the rod photoreceptors. Schematic representation of the GARP2 transgene. (A) tg cDNA driven by a 4.8 kb mouse opsin gene promoter, with a c-myc tag added near the C-terminus of GARP2 and mouse protamine polyA signal. The relative locations of the region of the transgene amplified by PCR for genotyping (679 bp) and RT-PCR (387 bp) are denoted by double headed arrows. (B) Reverse transcription of transgene-specific transcripts in tg retina. Total RNA was reverse transcribed and PCR amplified with a primer pair specific for the tg and the myc tag insertion sequence. The expected 387 bp PCR product was only observed in mice carrying the tg (Tg/+) and a control template sample (+control), but was not amplified from genomic DNA (genomic), in the absence of reverse transcriptase (no-RT) or in the absence of template DNA (-template). (C) Western blot with a myc tag antibody showing tg protein expression in the retinas of 5 transgenic lines and absence in WT. Relative levels of ROS proteins in GARP2 transgenic mice. (D) Quantitation of ROS protein levels normalized to 100% WT. (E) Representative Western blots showing relative levels of ROS proteins PDE6G (n = 3), PDE6A (n = 3), PER2 (n = 4), GC1 (n = 3), CNGA1 (n = 4), CNGB1 (n = 3) and GARP2 (n = 6) in tg compared to WT. CNGA1 was increased 30% (P < 0.01). Total GARP2 expression was increased 3-fold (P < 0.001), PDE6G was elevated 1.4-fold, and other ROS proteins were unchanged.