Rpmi 1640 medium

Conditionally immortalized mouse glomerular podocytes were procured from Chinese cell line infrastructure resources. RPMI 1640 medium (Genview, USA) enriched with 10% FBS (Sciencell, USA), 100 U/ml penicillin, 100 μg/ml streptomycin and IFN-γ (10 to 20 U/ml) (Bioss, China) was used to culture the cells at 33°C in a 5% CO₂ atmosphere. When the podocytes reached 80% confluency, the cells were passaged and stimulated to differentiate at 37°C in the absence of IFN-γ over a period of over seven days. Before proceeding to other experiments, the cells were maintained in a serum-free medium for 24 hours.
The cells were divided into the following groups: control cohort (11.1 mmol/L glucose); high glucose (HG) cohort (30 mmol/L glucose); miR-155 mimic or inhibitor cohort (50 nM miR-155 mimic or inhibitor); mimic or inhibitor normal control cohort (50 nM miR-155 mimic/inhibitor negative control); high glucose + miR-155 mimic/inhibitor or mimic/inhibitor control cohort; high glucose +miR-155 inhibitor + SIRT1 siRNA or control cohort (50 nM siSIRT1 or siRNA negative control). Cells in all groups were incubated with these reagents for 48 hours.

Human colon cancer DLD-1 cells were purchased from the Cell Bank of the Chinese Academy of Sciences (catalog number TCHu134). After resuscitation, DLD-1 cells were cultured in RPMI-1640 medium (Genview, USA) containing 10% heat-inactivated fatal bovine serum (Gibco, USA) and 1% antibodies (100 U/mL penicillin, 100 mg/mL streptomycin) in a 37°C, 5% CO₂ saturated humidity incubator (Zhou et al., 2018b (link)).