Restoration of decreased phosphorylation of GSK-3β by the treatment of TDZD-8 in the hearts of Ogt-Tg mice after TAC4W. (A) Western blot analysis for OGT, O-GlcNAc, ANP, Col III, and phosphorylation of GSK-3β, Smad2 in heart tissues from TAC4W-induced WT and Ogt-Tg mice with or without GSK-3β inhibitor (TDZD-8 (10 mg/kg/day, 3 weeks, IP)) treatment. Representative data was designated. (B–G) Quantifications of OGT, O-GlcNAc, ANP, Col III, and the ratios of p-GSK-3β/GSK-3β, p-Smad2/Smad2 expression intensity in (A) from three independent experiments using ImageJ software. The data were evaluated by two-way analysis of variance (ANOVA) followed by Tukey’s test. Values are shown as mean ± SD. *P<0.05. (H) Immunoprecipitation for O-GlcNAcylated GSK-3β in heart tissues from TAC4W-induced WT and Ogt-Tg mice with or without GSK-3β inhibitor (TDZD-8 (10 mg/kg/day, 3 weeks, IP)) treatment. Representative data was designated. (I) The intensity of each band for the O-GlcNAcylated GSK-3β expression (n=3) was measured using ImageJ software, and evaluated by two-way analysis of variance (ANOVA) followed by Tukey’s test. Values are shown as mean ± SD (n=3). *P<0.05. TAC, transverse aortic constriction; ANP, atrial natriuretic peptides; Col III, collagen type III.