Scrambled sirna and target gene specific sirnas

BV2 microglial cells were provided by Shanghai Institute of Materia Medica (Shanghai, China) and cultured in DMEM supplemented with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin in a humidified 5% CO₂–95% air environment at 37°C.
BV2 cells were seeded at the density of 30 × 10⁴ cells per well in six‐well plates and transfected with siRNAs (10, 100, and 200 nm) targeting PPARγ using Lipofectamine^® 2000 reagent (Invitrogen, NY, USA) according to the manufacturer's instructions (sequences are listed in Table 3). More than 90% knockdown of the targeted proteins was observed after 500 nm siRNA treatment. Scrambled siRNA and target gene‐specific siRNAs were purchased from GenePharma (Shanghai, China).

Primary microglia were seeded at the density of 120 × 10⁴ cells per well in six‐well plates and transfected with siRNAs (50 nm, 200 nm, and 500 nm) targeting LKB1 using Lipofectamine^® MessengerMAX mRNA Transfection Reagent (Invitrogen, NY, USA) according to the manufacturer's instructions (sequences are listed in Table 2). More than 90% knockdown of the targeted proteins was observed after 500 nm siRNA treatment. Scrambled siRNA and target gene‐specific siRNAs were purchased from GenePharma (Shanghai, China).

Astrocytes were seeded at the density of 10⁴ cells per well in six-well plates and transfected with siRNAs (50 nM, 200 nM, and 500 nM) targeting UCP2 using Lipofectamine Messenger MAX mRNA Transfection Reagent (Invitrogen, New York, NY, USA) according to the manufacturer’s instructions (sequences are listed in Table 2). More than 50% knockdown of the targeted proteins was observed after 500 nM siRNA treatment. Scrambled siRNA and target gene-specific siRNAs were purchased from Gene Pharma (Shanghai, China).