Silencer select small interfering rna sirna

All in vitro experiments were performed using the Sh-SY5Y (SY5Y) cells between 4–16 passages from the ATCC. Cells were grown in high glucose Dulbecco´s Modified Eagle Medium (DMEM) (Gibco, Grand Island, NY, USA) supplemented with 10% Fetal Bovine Serum (FBS) (Gibco) and 1% Penicillin-Streptomycin (PS) (Sigma, St. Louis, MO, USA). Cell lines were maintained at 37 °C with 5% CO₂. Repeated depolarizations were performed using 100mM potassium chloride (KCl) in DMEM added to the SY5Y cells for 5 min and then replaced with DMEM every two hours to create a sustained, activated MAPK signaling profile, as previously described [12 (link)]. MAPK inhibition was achieved using the MEK inhibitor PD184352 (PD18) at a final concentration of 10mM in DMSO.
For transfections, cells were electroporated using the Neon electroporation apparatus using the following parameters: 1200 V, pulse number 2, pulse width 20. Said parameters were based on previously described settings in the SY5Y cell line [12 (link)]. Silencer® select small interfering RNA (siRNA) targeting the lncRNAs BC028229, AK023739, CR615000, BC018494, and control siRNA were purchased from ThermoFisher and used at a final concentration of 100nM.