Si 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.

The sequences of siRNA targeting lncRNA MEG3 and scrambled negative control siRNA (si-NC) were designed and synthesized by GenePharma (Shanghai GenePharma Co., Ltd., Shanghai, China) according to the previously published protocol (15 (link)). pcDNA-MEG3 and empty vector (pcDNA) were synthesized by GenePharma (Shanghai GenePharma Co., Ltd.). At 60% confluence, HA-VSMCs were cultured in Dulbecco’s modified Eagle’s medium (Gibco; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO₂ for 24 h. HA-VSMCs were transfected with si-NC, si-MEG3 at a final concentration of 20 nmol/l in accordance with the manufacturer’s protocol. Furthermore, HA-VSMCs were transfected with pcDNA-MEG3 at a final concentration of 2 mg/l using. The following sequences were used: si-lncRNA MEG3, 5′-GGGCTTCTGGAATGAGCAT-3′; si-NC, 5′-UUCUCCGAACGUGUCACGUTT-3′ (15 (link)). HA-VSMCs were transfected using Lipofectamine^® 2000 on 6-well plates (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer’s protocol. Cells were harvested for the subsequent experiments 24 h after transfection.