Journal: PLoS ONE
Article Title: Defects in the Outer Limiting Membrane Are Associated with Rosette Development in the Nrl −/− Retina
doi: 10.1371/journal.pone.0032484
Figure Lengend Snippet: To analyze the integrity of the OLM in the Nrl −/− retina, sections were stained with antibodies specific for Zo-1 and beta-catenin (red), components of the OLM junctions, and S-opsin (green) to label OSs. A. In the WT retina, the Zo-1 staining forms a straight continuous line. In the Nrl −/− retina, gaps in OLM staining (arrows) can be seen that correspond spatially and temporally with the appearance of nascent rosettes and aberrant photoreceptors (arrowheads). B. Beta-catenin staining can be seen at the OLM as well as in the OPL and INL. In the Nrl −/− retina, gaps in OLM staining (arrows) can be seen that correspond with the appearance of nascent rosettes and aberrant photoreceptors (arrowheads). OLM: outer limiting membrane, ONL: outer nuclear layer, INL: inner nuclear layer. Scale bar, 20 µm.
Article Snippet: Primary antibodies were used as follows: rabbit anti-S-opsin (generated in-house and characterized previously ) diluted at 1∶1000; goat anti-S-opsin (H-17, Santa Cruz Biotechnology, Santa Cruz, CA) diluted at 1∶500; rabbit anti-recoverin (generously shared by Dr. James F. McGinnis, OUHSC ) diluted at 1∶5000; mouse monoclonal anti-BrdU (G3G4, the monoclonal antibody developed by Stephen J. Kaufman was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biology, Iowa City, IA) diluted at 1∶10; rabbit anti-B-catenin (C2206, Sigma-Aldrich, St. Louis, MO), diluted at 1∶1000; rabbit anti-ZO-1(Mid) (40–2200, Invitrogen, Carlsbad, CA) diluted at 1∶100; mouse monoclonal anti-ezrin (3C12, Abcam, Cambridge, MA) diluted at 1∶50; rabbit anti-mCAR-LUMIj (generously shared by Dr. Cheryl Craft, USC , , , ); rabbit anti-water channel aquaporin 4 (A5971, Sigma-Aldrich, St. Louis, MO) and rabbit anti-RDS-CT (generated in-house and characterized previously ) diluted at 1∶2000.
Techniques: Staining