Fig. 5.

4E-BP1/2 is necessary for diabetes-induced Cd40 mRNA translation in retina. Diabetes was induced in WT and 4E-BP1/2 double knockout (4EBP-DKO) male mice using STZ. Nondiabetic controls were administered vehicle (Veh). The retinas were collected after 3 months of diabetes. A, blood glucose concentrations were evaluated. B, protein O-GlcNAcylation and expression of 4E-BP1 and tubulin were assessed by Western blotting of retinal lysates. Protein molecular mass (in kDa) is indicated at the right. C, Cd40 mRNA expression was evaluated in retinal lysates by PCR. D, ribosomes from retinas were separated into either heavy or light fractions as described in Fig. 1. The distribution of the Cd40 mRNA was assessed in each fraction by PCR. E, CD40 and tubulin protein expression were assessed in retinal lysates by Western blotting. F, Nos2 mRNA abundance was evaluated in whole retina lysates by PCR. The results are expressed as the relative percentage of the mRNA in the polysomal fraction. The values are means ± S.D. The data were analyzed by two-way ANOVA (Table S1). Dunnett's post hoc analysis was used to identify differences between means when significant main or interaction effects were detected. *, p < 0.05 versus vehicle; #, p < 0.05 versus WT.