Figure 4

Immunohistochemical localization of the cleaved forms of caspase-3 (A) and caspase-9 (B) to retinal ganglion cells in retinas from untreated mice with 3 wks of intraocular hypertension. Caspases (green), NeuN (red), 4′,6-diamidino-2-phenylindole (DAPI) nuclear counterstain (blue) and merged images (yellow) are shown for each caspase, in the experimental glaucoma eye (GL) and in the fellow eye from untreated (NT) mice, and from mice with RHP before intraocular hypertension. Arrowheads in the GL eye of the NT group show immunopositive cleaved caspase-3 and -9 cells that colocalize with NeuN immunopositive cells. GCL, ganglion cell layer; INL, inner nuclear layer; IPL, inner plexiform layer. Scale bar in each = 20 µm. (C) Western blot results for cleaved caspase-9 protein. On the left are quantified results in histogram form (normalized to sham controls) and on the right are representative blots from each experimental group, using β-actin as a loading control. *P < 0.05 versus the corresponding eye in the control group or NT group. #P < 0.05 versus the NT group.