Mutant retinae contain excess numbers of amacrine and bipolar cells as well as Müller glia. (A) Representative images of antibody stainings against HuC/D (yellow), a marker for amacrine and ganglion cells, show HuC/D expression in the INL and GCL of both mutant and wildtype retinae. Nuclei are stained with DAPI (white). (B) Quantification of the overall number of HuC/D positive cells in both INL and GCL revealed no statistical difference. (C, D) Quantifications of HuC/D positive cells in individual layers indicated a significant reduction in the GCL (C) and a significant increase in the INL of mutant retinae (D). (E) Representative images of bipolar cells labeled with PKCα (yellow) showed that cell bodies are located in the INL and GCL of both mutant and wildtype retinae. Nuclei were stained with DAPI (white). (G) Quantification of PKCα positive cells showed a significant increase in mutant retinae. (F) Representative images of an antibody staining against the Müller glia cell specific marker GFAP (yellow) shows cell bodies of Müller glia cells that were located in the inner nuclear layer (INL) of wildtype and mutants’ retinae as well as their processes that span the entire retina. Nuclei were stained with DAPI (white). (H) Quantification of zrf1-positive cells revealed a significant increase in mutant retinae. Scale bars represent 25 µm. Statistics: all data are represented as mean ± SD, unpaired t-test, *P < 0.05; **P < 0.01; ***P < 0.001, or ****P < 0.0001. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer.
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