Nc si nc

Cells were placed into 6-well plates until they reached 70-80% confluence. Small interfering (si)RNAs targeting HOXC13 (si-HOXC13#1 and si-HOXC13#2) and the corresponding negative control (NC; si-NC) were purchased from GeneChem, Inc.; the sequences were as follows: si-HOXC13#1 sense, 5'-UUAACAUUAAAUACUCUUCUG-3' and antisense, 5'-GAAGAGUAUUUAAUGUUAAGG-3'; si-HOXC13#2 sense, 5'-UCCUUAACAUUAAAUACUCUU-3' and antisense, 5'-GAGUAUUUAAUGUUAAGGAAA-3' and si-NC sense, 5'-UUCUCCGAACGUGUCACGUTT-3' and antisense, 5'-ACGUGACACGUUCGGAGAATT-3'. pcDNA3.1 vectors containing full-length HOXC13 gene were used and empty pcDNA3.1 vectors (Shanghai GenePharma, Co., Ltd.) were used as the negative control. pcDNA3.1 containing HOXC13 (Oe-HOXC13) and DNMT3A (Oe-DNMT3A) and the corresponding NC plasmid (Oe-NC) were synthesized by Shanghai GenePharma, Co., Ltd. Transfection with aforementioned plasmids and siRNAs at a final concentration of 10 nM was performed using Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) at 37˚C for 48 h, according to the manufacturer's instructions and the transfection was completed. Further experiments were performed after 48 h.

Two small interfering (si)RNAs specific to SPTBN2 (siRNA-SPTBN2-1, 5'-ACGTCAATGTACACAACTTCACC-3'; and siRNA-SPTBN2-2, 5'-ACCATTACTTCTCCAAGATGAAG-3'), a pcDNA3.1 expression vector carrying ITGB4 (Ov-ITGB4) as well as their corresponding negative controls (NC) si-NC (5'-AAGACAUUGUGUGUCCGCCTT-3') and Ov-NC (empty vector) were purchased from GeneChem, Inc. Using Lipofectamine^® 2000 (Thermo Fisher Scientific, Inc.), 100 nM siRNA-SPTBN2-1/2, siRNA-NC and 4 µg Ov-ITGB4 and Ov-NC were transfected into A2780 cells at 37˚C for 24 h, according to the manufacturer's protocol. After 48 h, cells were washed with PBS and incubated in DMEM and reverse transcription-quantitative PCR (RT-qPCR) and western blotting were performed to test transfection efficacy.