Fig. 2

Prpf31 protein and its mRNA are highly expressed in the RPE of mouse retinas. Immunohistochemical staining showed strong Prpf31 signal in the RPE of CD-1 mouse retinas (a; arrowheads). Anti-Rhodopsin antibodies were used as positive control for immunohistochemical staining (b). A negative control without primary antibodies is also present (c). Western blot (d) and qPCR (e) analysis of Prpf31 protein and mRNA expression in the neuroretina and RPE samples showed that it is mainly expressed in the RPE (d, e). Anti-Rhodopsin and anti-Rpe65 antibodies were used as controls for the neuroretina/RPE tissue fractions, and anti-Gapdh antibody was used as a loading control (d). For qPCR, Recoverin (f) and Rpe65 (g) mRNA expression levels were used as controls for the two different tissue fractions. The bars in the graphs e-g represent means of fold change ± SEM (n = 4 replicates of 3 samples in each group). Statistically significant differences were determined by t-test (*p < 0.05, **p < 0.01). RPE = retinal pigment epithelium, OS = outer segment, IS = inner segment, ONL = outer nuclear layer, INL = inner nuclear layer, GCL = ganglion cell layer. Scale bars represent 50 μm