Pre mir mirna negative non targeting control 1

Cell lines were transfected using the nucleofector II system (Lonza, Allendale, NJ, USA) with the following programs: C-16 for H929 and JJN3, G-16 for MM1S, and X-005 for U266. Cells were transfected with on-TARGET plus™ control pool or on-TARGET plus SMART pool Human DEPTOR (Dharmacon, Lafayette, CO, USA); pre-miR™ miRNA precursors pre-miR-135b, pre-miR-642a, and pre-miR™ miRNA negative non-targeting control#1 (Ambion, Austin, TX, USA); and microRNA inhibitors, hsa-miR-135b-5p miRCURY LNA™ microRNA inhibitor, hsa-miR-642a-5p miRCURY LNA™ microRNA inhibitor, and miRCURY LNA™ microRNA inhibitor negative control A (Exiqon, Woburn, MA, USA). Small interfering RNA (siRNA) and miRNA concentration of 25 nM was used in all the experiments.

H929 and MM1S cell lines were transfected with Pre-miR™ miRNA precursors pre-miR-223 or pre-miR™ miRNA-negative, non-targeting control#1 (Ambion) at 50 nM concentration, using the nucleofector II system with C-16 program and Q-023 program, respectively (Amaxa). Transfection efficiency was assessed with Block-iT™ Fluorescent Oligo (Invitrogen) by flow cytometry.