siRNA knockdown in cells

IMR-32 and SK-N-AS cells were trypsinized and reverse transfected using Lipofectamine® RNAiMax (Life Technologies) and 50 μM of the following siRNAs (Life Technologies): GAS5 Silencer® Select siRNA (cat #: n272331), TP53 Silencer® Select siRNA (cat #: s607), BRCA1 Silencer® Select siRNA (cat #: s459), GADD45A Silencer® Select siRNA (cat #: s225791), and Silencer® Select negative control #1 (cat #: 4390843). Cells were seeded into 6-well plates at a density of 2.5 × 10⁵ cells per well. Cells were incubated for 24 or 48 hours at 37°C in a CO₂ incubator. Once the transfection period was complete, cells were harvested and counted for use in cell biology assays. The efficiency of gene knockdown was assessed by quantitative real-time PCR (qRT-PCR).