Sirna construction kit

The activity of caspase-3 and PARP in HeLa, HEC-1-A, and KLE cancer cells were developed as previously reported (Zhang and Snyder, 1992 (link); Kang et al., 2012 (link)). Various siRNA sequences from the LKB1 RNA sequence were prepared utilizing a siRNA Construction kit (Ambion, Austin, TX, USA). After selection of two clones, the prepared sequences were then transfected with oligofectamine (Thermo Fisher Scientific, Carlsbad, CA, USA) in accordance with supplier’s recommendation. TRIZOL Reagent (Thermo Fisher Scientific) was used to isolate whole RNA and reverse transcription (RT) PCR was then conducted. LKB1 expression was measured via western blot analysis.

Anti mouse and anti rabbit IgG-horse radish peroxidase conjugate antibodies were purchased from Pierce Company. An ECL (enhanced chemiluminiscence) kit was purchased from Amersham. Lipofectamin, Plus^™ reagent, Trizol and reverse trancriptase (Superscript II RT) were purchased from Invitrogen (USA). Bioneer (Korea) synthesized all primers and miR-inhibitor used in this study. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and all other chemicals used in this study were purchased from Sigma. Anchorage-independent growth assays kit was purchased from Millipore. Annexin V-FITC kit for the detection of apoptosis was purchased from Biovision. Transwell chamber system with 8-μm pore polycarbonate filter inserts for the determination of invasion potential was purchased from CoSTAR (USA). Quick-change site-directed mutagenesis kit was purchased from Stratagene. miRNA array was purchased from Signosis Company. SiRNA construction kit, mirVana miRNA isolation kit and SYBR Green qRT-PCR kit were purchased from Ambion (USA). qScript™ reverse transcriptase was purchased from Quanta Biogenesis. A-Plus Poly(A) Polymerase Tailing Kit was purchased from Cell Script. SYBR PCR Master Mix was purchased from Applied Bosystems.