Fig. 5
Hspa4l is highly expressed in the RPE of Prpf31A216P/+ mice. Analysis of Hspa4l expression by qPCR in the neuroretina and RPE samples show that Hspa4l mRNA is overexpressed in the RPE of Prpf31A216P/+ mice (a). The boxplot a represents the fold change of Hspa4l expression in the neuroretina and RPE of WT and Prpf31A216P/+ mice (WT n = 3 and Prpf31A216P/+ n = 6). Statistically significant differences were determined by Mann-Whitney U-test (*p < 0.05). Western blot of RPE samples showed higher expression of Hspa4l protein in Prpf31A216P/+ mice compared to WT (b). Anti-Gapdh antibodies were used as loading control (b). Whole-mount of the RPE obtained from WT (c-h) and Prpf31A216P/+ mice (i-n) were immunostained with anti-Prpf31 (c, f, i, l) and anti-Hspa4l antibodies (d, g, j, m). TRITC-phalloidin was used to stain F-actin microfilaments (c-n; blue). Magnified images are (f-h and l-n) and merged are shown (e, h, k, n) Prpf31 signal was mainly distributed in the nuclei of RPE cells in WT mice (c, f; arrowhead), while Prpf31 protein aggregates were observed in the cytoplasm (i, l) colocalizing with Hspa4l signal in mutant Prpf31A216P/+ mice as shown in the merged images (k, n). Prpf31 signal was very low in the nuclei of Prpf31A216P/+ mice (l; arrow). Scale bars represent 25 μm