Fig. S5.

Accumulation of methylglyoxal-derived CEL but not glyoxal-derived CML in HG retinas. (A) The CEL concentration is linearly related to the retinal damage score. (B) Immunofluorescent detection of CEL using an antibody independent of that shown in Fig. 4 E and F confirms that CEL levels are high in HG retina compared with LG or HGxoLG retina. (Left) Staining in the neural retina was largely confined to the inner nuclear layer (INL) with distinct processes that terminate at the outer and inner limiting membranes. This pattern resembles staining within Müller glial cells, which have been reported to express high levels of CEL in diabetic rats. (Right) CEL was expressed at high levels throughout the HG RPE but only weakly in LG and HGxoLG RPE. The white line indicates the separation between the RPE layer and choroid, which shows high background because of anti-mouse secondary antibodies. (C) Boxplot of CML levels in LG and HG plasma show that CM) levels are not different in the dietary groups; n.s, not significant. (D) Immunohistochemical detection of CML shows that it is broadly detected throughout the retinas of mice from all dietary groups. Sample size in C is n = 8. (Scale bars: 100 μm in B, Bottom Left, and D; 50 μm in B, Bottom Right.)