The short hairpin (sh)RNAs specifically targeting LINC00491 and a negative control shRNA (sh-NC) were designed and synthesized by Shanghai GenePharma Co., Ltd. KYSE30 and KYSE410 cells in the logarithmic growth phase were seeded in a petri dish. Cell transfection with shRNA was performed using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to manufacturer's protocol; the ratio of shRNA (500 µg/µl): Lipofectamine® 2000 (µl) was 1:3. The sequences of shRNAs were as follows: sh-LINC00491#1, 5′-GGT GTA TTC CAC ATT GTC TCT-3′; sh-LINC00491#2, 5′-GGC CAA AGG TCT GAT AAT TGC-3′; sh-LINC00491#3, 5′-GGA TAT GTG CAG GGA GTC TAG-3′; and sh-NC, 5′-TTC TCC GAA CGT GTC ACG T-3′.

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