Si rna meg 3

All siRNAs were obtained from Invitrogen; Thermo Fisher Scientific, Inc. The following siRNAs were used: si-RNA-MEG-3 (si-MEG-3; sense: 5′-CAUUGGCAUCCUUCGAAAUTT-3′ and antisense: 5′-AUUUCGAUGGAAGCCAAUGTT-3′) or si-RNA-vector (sense: 5′-AAGCUGAGCAAGAUUCAGACC-3′, and antisense: 5′-GGUCUGAAUCUUGCUCAGCUU-3′; Invitrogen; Thermo Fisher Scientific, Inc.). In brief, BV-2 and BC3H1 cells (1×10⁶) were transfected with 100 pmol si-MEG-3 or si-RNA-vector using Cell Line Nucleofector kit L (Lonza Group, Ltd., Basel, Switzerland). After 72 h transfection, si-RNA-MEG-3-transfected BV-2 and BC3H1 cells (1×10⁶) were treated by tunicamycin (2 mg/ml, si-MEG-3-TUN) for 12 h at 37°C.

All siRNAs were synthesized by Applied Biosystems; Thermo Fisher Scientific, Inc. including siRNA-MEG-3 (si-MEG-3, sense, 5′-CGAUUGGAGCGAUCAAGCUTT-3′ and anti-sense, 5′-AGCUUGAUCGCUCCAAUCGTT-3′) and siRNA-vector (sense, 5′-UUCUCCGAACGUGUCACGUTT-3′ and anti-sense, 5′-ACGUGACACGUUCGGAGAATT-3′). BV-2 and BC3H1 cells (1×10⁶) were transfected with 100 pmol of si-MEG-3 (Applied Biosystems) or siRNA-vector as a control (Applied Biosystems) using a Cell Line Nucleofector kit L (Lonza Group, Ltd., Basel, Switzerland). Further analysis was performed 48 h following transfection.