Si hottip

Si-HOTTIP (siRNA against HOTTIP, 5′-AGGCTGAGCTAATACAGTA-3′), si-NC, miR-92a-2 mimics, miR-NC, and si–c-Met (siRNA against c-Met, 5′-ATCTTGAGCCATTCACCGGAA-3′) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China) and transfected into mouse cardiomyocytes using Lipofectamine 2000 (Invitrogen). To overexpress HOTTIP and c-Met, the cDNA sequence of HOTTIP and c-Met were amplified with mouse genome as the template and cloned into pcDNA3.1 Expression vector to generate pc-HOTTIP and pc-c-Met. Cells transfected with the empty vector were used as the negative control (pc-NC). Similarly, pc-HOTTIP, pc-c-Met, or pc-NC was transfected into mouse cardiomyocytes using Lipofectamine 2000.

For gene interference studies, plasmids (si‐HOTTIP, si‐DNMT1, si‐SP‐C, oe‐HOTTIP, and their negative control) were purchased from Shanghai GenePharma Co. Ltd., and were transfected into cells using Lipofectamine^TM 3000 Transfection Reagent (L3000075, Invitrogen, Thermo). After a 1‐day incubation period, the transfection efficiency of plasmids was assessed using quantitative real‐time polymerase chain reaction (qRT‐PCR) and western blot analyses.