Sicon

Predesigned Src-targeting siRNA (siSrc) and control siRNA (siCon) were purchased from Santa Cruz. The cells were transfected with 100 nM siRNA while using Lipofectamine RNAiMAX (Invitrogen, Waltham, MA, USA) for 5 h, cultured in medium containing 0.5% FBS, and harvested ~48 h after transfection.

Small interfering RNA (siRNA)-targeting human apelin (si-apelin) and nonspecific siRNA (si-Con) were purchased from Santa Cruz Biotechnology. Transfection was performed using a Lipofectamine reagent (Invitrogen) according to the manufacturer’s instructions. At 24 h posttransfection, LX-2 cells were harvested and used for qRT-PCR and Western blot assays.

The human OS cell line 143B was purchased from ZhongQiaoXinZhou Biotechnology (NO.ZQ0455). The cells were routinely cultured in complete culture medium (ZhongQiaoXinZhou, NO.ZQ‐303) at 37°C in a 5% CO₂ atmosphere.
Transfection was performed at a cell density of 60% in a six‐well plate with small interfering (si) RNA targeting MCAM (si‐MCAM) or non‐specific control siRNA (si‐con; Santa Cruz Biotechnology; NO.sc‐35918 and sc‐37007, respectively) using Lipofectamine 2000 transfection reagent (Invitrogen; Thermo Fisher Scientific; NO.11668027) for 48 h, following the manufacturer's protocol. Thereafter, the effect of siRNA transfection was examined using western blot analysis, and the cells were used for subsequent experiments.