Sirnagdf11

Small interfering RNA (siRNA) specific for GDF11 gene expression (siRNA^GDF11) and control scrambled siRNA were synthesized by GenePharma Co., Ltd (Shanghai, China). Cells were seeded at 1x10⁵ per cell into 12‐well plates one day before transfection. Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) (2 μL/mL) was mixed with siRNAs (final 50 nmol/L) in reduced serum medium OptiMEM (Invitrogen) and then added into each well (1ml per well). Medium was changed to DMEM containing 10% FBS 8 hours after transfection and cultured for further analysis. Expression of GDF11 at levels of mRNA and protein was detected by RT‐PCR and Western blot, respectively. For the experiment of differentiation that lasted 14 days, the cells were transfected with siRNA again 7 days after the first transfection to maintain GDF11 being knocked down. VEGF165 was added to the culture 24 hours after either transfection to induce differentiation of MSCs to ECs.

Small interfering RNA (siRNA) specific for GDF11 gene expression (siRNA^GDF11) and control scrambled siRNA were synthesized by GenePharma Co., Ltd (Shanghai, China). Cells were seeded at 1 × 10⁵ cells per well into 12-well plates one day before transfection. The siRNAs (final 50 nM) were mixed with Lipofectamine 2000 (Invitrogen, USA) (2 μl/ml) in medium OptiMEM (Invitrogen, USA) and then added into each well (1 ml per well). Medium was changed into DMEM containing 10% FBS 8 h after transfection and cultured for further analysis. Expression of GDF11 at levels of mRNA and protein was detected by RT-PCR and Western blot, respectively.