From: Blocking Endothelin-B Receptors Rescues Retinal Ganglion Cells from Optic Nerve Injury through Suppression of Neuroinflammation Invest. Ophthalmol. Vis. Sci.. 2012;53(7):3490-3500. doi:10.1167/iovs.11-9415 Figure Legend: Immunohistochemistry for ETs, GFAP, and ETB receptors in the optic nerves of the sham controls and experimental animals with a crushed optic nerve. Representative pictures from three independent samples for each condition are presented. (A) Changes in the expression of ETs on days 7 and 14 after crushing the optic nerve. The sections were stained with rabbit anti-ETs and Alexa 594–conjugated goat anti-rabbit IgG secondary antibodies. Immunoreactivity to ETs (red) is expressed mainly at the crushed site on day 7 (arrows), which spreads and intensifies on day 14 (crush [PBS]) compared with the sham controls. Pretreatment with BQ-788 suppresses the increased immunoreactivity to ETs (crush [BQ-788]; crush site is indicated by arrows). Pretreatment with BQ-123 also suppresses the increase but the effects are less intense than those by BQ-788 (crush [BQ-123]). A total of 24 rats were used in this analysis. (B) Higher magnification view of the crushed site (area B) stained with antibodies to rabbit anti-ETs and mouse monoclonal anti-GFAP. Cells that have infiltrated into the crushed site are stained by DAPI (blue) and are also positively stained by antibodies to ETs (Alexa 594). Merged image by Photoshop indicates high incidence of colocalization of nuclear staining with DAPI and ETs (left panels). At the crushed site where a dense cellular infiltration with the expression of ETs can be seen, GFAP immunoreactivity is very weak. In the area surrounding the crushed site, immunoreactivity to GFAP (FITC) is increased as shown in Figure 2B (right panels). (C) Higher-magnification view of the crushed site (area C) stained with antibodies to sheep anti-ETB receptors and rabbit anti-ETs. Some cells that have infiltrated the crushed site are positively stained with both antibodies to ETB receptors (Alexa 488-conjugated donkey anti-sheep IgG) and ETs (Alexa 594-conjugated goat anti-rabbit IgG). (D) Double labeling of other crushed optic nerve sections stained with antibodies to CD68 and ETs. The migratory cells that are concentrated at the crushed site (arrows) are positively stained with CD68 (green) at high density. Some of these CD68-positive cells at the crushed site are also stained with ETs (red). Bar = 100 μm. Date of download: 11/8/2017 The Association for Research in Vision and Ophthalmology Copyright © 2017. All rights reserved.