Sigenome risc free

Transient knockdown of S100A11 and E-cadherin in MCF-10A cells was performed by transfection of MISSION Predesigned (Sigma) or custom-made siRNAs, respectively. To knockdown S100A11, the following siRNAs were used: SASI_Hs01_00164495 (si-S100A11#1) and SASI_Hs01_00164498 (si-S100A11#2). To knockdown E-cadherin, the following siRNAs were used: si-E-cadherin#1 (sense, 5′-CAUCUUGACUAGGUAUUGUCU-3′; antisense, 5′-AGACAAUACCUAGUCAAGAUG-3′) and si-E-cadherin#2 (sense, 5′-GAGAGAGUUUCCCUACGUAUA-3′; anti-sense, 5′-UAUACGUAGGGAAACUCUCUC-3′). All experiments using siRNAs were carried out in the presence of siGENOME RISC-Free (Dharmacon), used as a negative control for siRNA experiments (si-Control). MCF-10A cells were transfected with a final concentration of 50 nM of siRNAs diluted in Opti-MEM Reduced Serum Medium (Gibco) using Lipofectamine RNAiMAX (Invitrogen), following the manufacturer's protocol. Transfected cells were incubated for 48 h or 72 h before they were live imaged, lysed, or fixed and immunoprocessed.