Cell transfection

We purchased pcDNA3.1-NEAT1 and pcDNA3.1-TPD52 plasmids and the pcDNA3.1-vector from Guangzhou RiboBio Co., Ltd . Small interfering RNA (siRNA) targeting NEAT1 (si-NEAT1-1, 5'-CTCCTTTTGGACTTTTCTCTAGG-3' and si-NEAT1-2, 5'-GTGATTGCATTGCAGATTACTAG-3'); TPD52 (si-TPD52-1, 5'-GAGTGAACAAAAGCTATCTCTAC-3' and si-TPD52-2, 5'-TTGAAGAAAAGGTCGAAAACTTA-3'); miR-218-5p mimics (5'-UUGUGCUUGAUCUAACCAUGU-3'); miR-218-5p inhibitor (5'-ACAUGGUUAGAUCAAGCACAA-3'); and their respective controls (mimics control, 5'-UUACUCGACACGUGUCAAGUUU-3'; and inhibitor control, 5'-CAGUACUUUUGUGUAGUACAA-3'), with unrelated sequences, were also synthesized by Guangzhou RiboBio Co., Ltd. Lipofectamine^® 3000 (Thermo Fisher Scientific) was used to transiently transfect overexpression plasmids (1,000 ng/6 cm dish) and siRNAs (30 nM) into cells, which were subsequently cultured in a humidified incubator for 36 hours. Experiments were performed 36 hours after transfection. Si-NEAT1-2 and si-TPD52-2 were selected, as they have higher interference efficiency than si-NEAT1-1 and si-TPD52-1, respectively.