Bmpr2

Validated select siRNA was used to knockdown the expression of XIAP, BMPR2, ALK3, and ALK6 (Life Technologies). The ID numbers for the siRNA are: XIAP (S1456), ALK3 (s281), ALK6 (s2042), and BMPR2 (s2044 and s2045). Silencer Select negative control siRNA (4390843) was used to evaluate selectivity. Silencer Select negative controls do not target any gene product and have no effect on cell proliferation or viability. Transfections of the siRNA were performed using Lipofectamine® RNAiMAX Reagent (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s protocol. Briefly, H1299 and A549 lung cancer cell lines were seeded for 24 h (h) up to 70–80% confluence at the time of transfection. 150 μl of Opti-MEM® Medium was used to dilute 9 μl of Lipofectamine® RNAiMAX Reagent, control siRNA and target siRNA. Diluted Lipofectamine® RNAiMAX Reagent was mixed with diluted siRNA in 1:1 ratio and incubated for 5 min (min) at room temperature to obtain the RNA-lipid complex. The cells were incubated in siRNA-lipid complexes for 24 h at 37 °C. After 24 h, the media was changed to fresh media and the transfected cells were used for further experiments. Cells were transfected with 30 nM ALK3, 20 nM ALK6, 30 nM XIAP, and 6 nM BMPR2.