S8588

After successfully inducing the diabetic mouse model, we administered the ACSS2 inhibitor (Selleck Chemicals, S8588) by gavage at a dose of 50 mg/kg (of mouse body weight) once a day for a follow-up course of 12 weeks. Control mice received a vehicle solution. The mice were divided into 3 groups: Ctrl (n = 6), DM (n = 6), and diabetes with ACSS2 inhibitor (DM + ACSS2 inhibitor) (n = 6). Body weight and fasting blood glucose concentration were monitored during the experimental period. The mice were humanely sacrificed at 12 weeks after STZ treatment.

Immortalized mouse podocytes were a gift from Ai Hua Zhang from the Children’s Hospital of Nanjing Medical University (Nanjing, China). Mouse podocytes were cultured in RPMI 1640 medium (Wisent) with 10% fetal bovine serum and 10 U/mL recombinant mouse γ-interferon (γ-IFN; PeproTech) at 33°C for proliferation. Podocytes were then transferred to nonpermissive conditions at 37°C without γ-IFN for at least 7 days for differentiation (40 (link)). Podocytes were incubated in RPMI 1640 with 10% FBS in the presence of 30 mmol/L d-glucose (MilliporeSigma) as the final concentration. The cells were exposed to 11 mmol/L d-glucose as the control. According to the procedure recommendation, ACSS2 siRNA transfection was performed with Lipofectamine 2000 from Thermo Fisher Scientific. ACSS2 inhibitor (Selleck Chemicals, S8588) or DMSO as control was added to podocytes before HG treatment. The sequences of control siRNA and ACSS2 siRNA are listed in Supplemental Table 3; https://doi.org/10.1172/jci.insight.165817DS1

Immortalized mouse podocytes were a gift from Ai Hua Zhang from the Children’s Hospital of Nanjing Medical University (Nanjing, China). Mouse podocytes were cultured in RPMI 1640 medium (Wisent) with 10% fetal bovine serum and 10 U/mL recombinant mouse γ-interferon (γ-IFN; PeproTech) at 33°C for proliferation. Podocytes were then transferred to nonpermissive conditions at 37°C without γ-IFN for at least 7 days for differentiation (40 (link)). Podocytes were incubated in RPMI 1640 with 10% FBS in the presence of 30 mmol/L d-glucose (MilliporeSigma) as the final concentration. The cells were exposed to 11 mmol/L d-glucose as the control. According to the procedure recommendation, ACSS2 siRNA transfection was performed with Lipofectamine 2000 from Thermo Fisher Scientific. ACSS2 inhibitor (Selleck Chemicals, S8588) or DMSO as control was added to podocytes before HG treatment. The sequences of control siRNA and ACSS2 siRNA are listed in Supplemental Table 3; https://doi.org/10.1172/jci.insight.165817DS1