Fig. 1.

REV-ERBα declines in aging RPE and sub-retinal deposits increase in Rev-erbα^−/−mice. (A) Immunostaining of cross-sections from Wild-type (WT) mice shows REV-ERBα antibody staining (red) in RPE nuclei, co-localized with nucleus marker DAPI (blue). Scale bar: 10 μm. (B–C) Western blot (B) and quantification (C) of REV-ERBα protein levels in the RPE isolated from 2–3, 4–5, 7–8 and 10–11 months old WT mice. Protein size: REV-ERBα (67 kDa), β-Actin (42 kDa). n = 3 mice/group. Quantification statistics was performed with nonparametric Kruskal-Wallis test. (D) Rev-erbα^−/− retinas show an increasing number of whitish abnormal deposits (yellow arrowheads) in fundus imaging with aging at 3, 6, and 12 months old (n = 10–12 eyes from 5 to 6 mice/group, mixed genders). (E) Quantification of the total lesion numbers in fundus images of Rev-erbα^+/+and Rev-erbα^−/− mice during aging (n = 10–12 eyes/group). (F) Rev-erbα^−/− retinas show oil red O-positive staining (red) as lipid marker in the sub-retinal deposits (red), which are counter-stained with hematoxylin in 5-month-old eyes; in 12-month-old eyes sub-retinal deposits are stained with oil red O only. Scale bar: 20 μm. Error bars indicate SD. *: P < 0.05; ****: P < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)