Human yap sirna sc 38637

Primary human pulmonary arterial smooth muscle cells (hPASMCs, c-12521) were purchased from American Type Culture Collection (Rockville, MD, USA) and cultured in Dulbecco's Modified Eagle's Medium (DMEM, 1101, ScienCell) supplemented with 2.5% fetal bovine serum (FBS), 1% penicillin/streptomycin (PS) solution and 1% smooth muscle cell growth supplement (SMCGS). All cells were grown at 37 °C in humidified air with 5% CO₂. Cells at passages 3–9 were used for the experiments.
For hypoxia (3% O₂) experiments, hPASMCs were cultured in serum-free DMEM for 12 h before treatments and then placed in a Heracell Vios 150i CO₂ incubator (Thermo Fisher Scientific) for 24 h. For the normoxia control, hPASMCs were cultured in normal incubators with 21% O₂ for 24 h.
Transfections were performed using Lipofectamine® 3000 Transfection Reagent (L3000075, Invitrogen, CA, USA) according to the manufacturers’ instructions. Short-interfering RNAs targeting SIK1 (SIK1 siRNA) (sense: 5′-CCACUUUGCUGCCAUUUAUTT-3′, antisense: 5′-AUAAAUGGCAGCAAAGUGGTT-3′) and a relative scrambled siRNA were designed and synthesized by GenePharma (China). Human YAP siRNA (sc-38637) was purchased from Santa Cruz Biotechnology. SIK1 adenovirus (Ad SIK1) and null adenovirus (Ad null) were purchased from GeneChem (China). Ad SIK1 was used to overexpress SIK1.