Anti mir 9

miR-9-mimics, anti-miR-9, NC-mimics, anti-NC, ABCC1 (ATP binding cassette subfamily C member 1), siRNA (si-ABCC1) and si-NC were designed and synthesized by RiBoBio. Lipofectamine 2000 was applied for transfection. The lentiviral empty vector (NC-LV) expressing GFP and GFP with high miR-9 (LV-miR-9-5p) were constructed by Systems Biosciences. Virus establishment and cell transduction were carried out in basement membrane cells, and cells were obtained using puromycin (1μg/mL) and sorted using flow cytometry to maintain at 95% of GFP positive rate.

miR-9-5p mimic control (miR-Con), miR-9-5p mimic (miR-9), miR-9-5p inhibitor control (anti-miR-Con), and miR-9-5p inhibitor (anti-miR-9) were synthesized by Ribobio (Guangzhou, China). Primary mouse microglia and BV-2 cells were seeded at 1 × 10⁵ cells per well in a 24-well plate in 1 ml of antibiotic-free standard growth medium and grown to 40–80% confluency. Then, cells were transfected with 50 nM miR-Con or miR-9 or with 100 nM anti-miR-Con or anti-miR-9. 1 μl RiboFECT™CP reagent (RiboBio, Guangzhou, China) was added to each well according to the manufacturer’s protocols. The medium containing RiboFECT™CP reagent was replaced after 24 h of transfection. Transfection efficiency was determined via western blotting.