2.5. Gene Downregulation

To study the physiological functions of MDL1AS, its expression was modulated with double-stranded interfering RNAs (DsiRNAs), as described [29]. All DsiRNAs were synthetized by Integrated DNA Technologies (Coralville, IA, USA) at 10 nmol scale (Table 1). They were diluted in RNAse-free water to a final concentration of 100 µM. For transfection, HCT116 (150,000–250,000 cells per well), MCF7, and MDA-MB-231 (50,000–100,000 cells per well) cells were seeded in six-well plates and transfected with Lipofectamine RNAiMAX (Thermo Fisher, Waltham, MA, USA), according to manufacturer’s instructions, in serum-free medium and using DsiRNA at a concentration of 10 nM.

Sequence of the DsiRNAs used to downregulate MDL1AS expression.