Plasmid and short hairpin RNA (shRNA) transfection

OVCAR3 and CAOV-3 cells were plated into a 6-well plate (1×10⁵ cells/well), cultured overnight, and were then transfected with 50 nM corresponding vectors using Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturers instructions. The shRNA targeting DDX3 (shDDX3), control shRNA (shCtrl), pcDNA-DDX3 and the control plasmid pcDNA3.1 were provided by Novogene Co., Ltd. The sequences of the targeted genes were as follows: shDDX3, F 5-AACCCACCACAGCUAGAACTT-3, R 5-AAG UUCUAGCUGUGGUGGGTT-3; shCtrl, F 5 UUCUCC GAACGUGUCACGUTT-3, R 5 ACGUGACACGUU CGGAGAATT-3; and pcDNA-DDX3, F 5 GAAGCT ACTAGAGGTTTCTAC-3 and R 5 TCTCAACATCAC TGAAACTTTC-3. Analyses were performed 48 h after transfection and replicated three times to produce the experimental data.