SMCs and VSMCs (ATCC, Manassas, VA) were maintained in DMEM supplemented by 10% of foetal bovine serum, 100 U/mL of penicillin, and 100 μg/mL of streptomycin (all cell culture reagents were acquired from Gibco, Thermo Fisher Scientific). The cells were maintained in a 37°C incubator under 5% CO2 and passaged every 3 days via digestion with 0.25% of trypsin. In this study, to study the effects of circORC2 on the expression levels of miR‐19a, NST mRNA, and MLN mRNA, SMCs and VSMCs were divided into 2 cell models, as shown below. In cell model 1, SMCs and VSMCs were divided into 2 groups, that is (a) Empty vector group (SMCs and VSMCs transfected with an empty vector) and (b) P‐circORC2 group (SMCs and VSMCs transfected with the vector carrying circORC2). In cell model 2, SMCs and VSMCs were also divided into 2 groups, that is (a) NC group (SMCs and VSMCs transfected with a negative control), and (b) CircORC2 siRNA group (SMCs and VSMCs transfected with circORC2 siRNA). In both cell models, SMCs and VSMCs were transfected with corresponding plasmids using Lipofectamine 2000 (Invitrogen) in accordance with the recommended transfection protocol shown in the manufacturer manual. At 48 hours after transfection, the cells were harvested for subsequent analyses.

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