Supplementary Data

Stylianos Michalakis, Heidi Geiger, Silke Haverkamp, Franz Hofmann, Andrea Gerstner, and Martin Biel

Files in this Data Supplement:

• Supplementary Table S1 - 23.3 KB

• Supplementary Figure S1 - 224 KB
  Specificity of anti-opsin antibodies. (A-D) Confocal images of wild-type retinal slices double-labeled with polyclonal anti-MWS or anti-SWS antibody (green) and the nuclear dye Hoechst (blue). Anti-MWS labels the majority of cone outer segments in the dorsal (A) but not in the ventral (B) mouse retina. By contrast, anti-SWS labels only a few cones in the dorsal retina (C) but the majority of cone outer segments in the ventral mouse retina (D). (E, F) Co-staining of a wild-type dorsal section with polyclonal anti-MWS opsin antibody (green), monoclonal COS1 antibody (red, specific for MWS cones) and PNA (blue). There is complete overlap between MWS and COS1 staining (yellow overlay in (E)). Triple staining shown in (F) reveals that all cones in this section are MWS-type cones. (G) Whole-mount preparation from the central retina stained with polyclonal anti-MWS antibody and the monoclonal OS2 antibody (specific for SWS-type cones). There is only sparse overlap of signals (yellow) confirming the specificity of the two antibodies. (H) Western blots with 30 µg retinal membranes from adult wild-type mice using polyclonal anti-MWS (left) or anti-SWS (right) detected a 42kDa and 46 kDa band, respectively. Scale bars: (A-D) 100 µm; (E) 20 µm; (G) 10 µm.

• Supplementary Figure S2 - 190 KB
  Preserved rod morphology in CNGA3-deficient mice. (A, B) Confocal images of retinal sections from the ventral part of PM17 wild-type (A) and CNGA3^-/- (B) retina labeled with anti-rhodopsin antibody (red) and PNA (green). (C-E) Normal expression of CNGA1 (C), CNGB1 (D) and GCF (E) in the retina of a 22-month-old CNGA3^-/- mouse. The confocal scans in (C, D) were overlaid with the respective differential interference contrast images to demonstrate the retinal morphology. Scale bar: 20 µm.