Fig. 3

sa0143 is a null mutation of optn with reduced protein and transcript levels.

A. Trace files from sequencing reactions of adult fish showing region of sa0143 mutation and corresponding amino acids. Arrow indicates mutated base. B, C. Live pictures of adult WT (B) and optn (C) fish showing no difference in morphology. D. Western blot of protein extracts from 3 dpf embryos. Green = anti-Optn, Red = anti-γ-tubulin as a loading control. Asterisks indicate likely non-specific bands. The Optn band is absent in the MZoptn lane. E. Representative gel of semi-quantitative RT-PCR run with cDNA from retinas of 6 mpf fish. Top blot was used with primers against exons 1-3 of optn, bottom blot was used with primers against ef1a as a loading control. PCR was stopped at cycle times indicated below the blots. F. Quantification of 3 semi-quantitative RT-PCR experiments. Levels were analyzed for 28-cycle bands and normalized to WT bands for each experiment. ** indicates p≤0.01.