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
Recombinant mouse γ interferon γ ifn
Manufactured by Thermo Fisher Scientific
Recombinant mouse γ-interferon (γ-IFN) is a protein produced using recombinant DNA technology. It is a cytokine that is involved in the modulation of immune responses.
Automatically generated - may contain errors
2 protocols using recombinant mouse γ interferon γ ifn
1
Differentiated Mouse Podocyte Culture
2
Differentiated Mouse Podocyte Culture
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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
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