Diabetes was induced by intraperitoneal injection of STZ. The corresponding control mice were treated with vehicle (Ctrl). Diabetic mice were treated with vehicle (DM) or ACSS2 inhibitor (50 mg/kg) (DM+ACSS2 inhibitor). The mice were euthanized at week 12. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 vs. Ctrl; ^#P < 0.05, ^##P < 0.01, ^###P < 0.01 vs. DM, n = 6). (A) Kidney hypertrophy was determined by the ratio of kidney weight to body weight (n = 6 biological replicates, 1-way ANOVA). (B) Urinary ACR was measured by commercial ELISA for microalbuminuria and creatinine detection assay (n = 6 biological replicates, 1-way ANOVA). (C) Representative images of PAS-stained kidney sections (original magnification ×400; scale bars, 100 μm). Bar graph analysis shows the quantification of the mesangial expansion area percentage (n = 10 biological replicates, 1-way ANOVA). (D) Representative images of glomerular ultrastructural change such as podocyte effacement and GBM thickness were observed by electron microscopy (original magnification ×40,000; scale bars, 500 nm). (E) Representative immunohistochemical staining images of α-SMA in kidney sections (original magnification ×200; scale bars, 200 μm). (F) Representative confocal microscopic images showing the expression of LC3 (green) and WT-1 (red). Nuclei were stained with DAPI (blue). The quantifications of WT-1 (green) per glomerulus in the kidney (n = 20 biological replicates, 1-way ANOVA) were analyzed (original magnification ×400; scale bars, 50 μm). (G) Representative confocal microscopic images showing the expression of p-S6K1 (green) and WT-1 (red). Nuclei were stained with DAPI (blue) (original magnification ×400; scale bars, 50 μm). (H) Western blotting analysis for raptor and the phosphorylation activation of mTOR, S6K1, 4EBP, and LC3 in renal cortex tissues (n = 3 biological replicates, 1-way ANOVA).