Figure 6. TWIST2 activates LINC01235 expression through a positive feedback loop. (A) RT-qPCR shows ectopic TWIST2 upregulated LINC01235 expression in HGC-27 and SGC-7901 cells. (B) Putative TWIST2-binding sites in the LINC01235 promoter. (C) Luciferase reporter assays confirmed TWIST2 activation of LINC01235 promoter through TWIST2-binding sites in HGC-27 and SGC-7901 cells. (D) ChIP assays were carried out using IgG and TWIST2 antibody in HGC-27 and SGC-7901 cells, followed by RT-qPCR with primers amplifying the LINC01235 promoter region. (E) Venn diagram indicates the miRNA acts as a sponge in three databases, including Lncbase, miRWalk and miRDB. (F, G) The correction of miR-6852-5p with LINC01235 and TWIST2 in TCGA database. (H) Schematic diagram shows the LINC01235–TWIST2–THBS2 axis; TWIST2, in turn, upregulates LINC01235. WT: wild type, and Del: deleted type. Results are expressed as means ± SD. **: P < 0.01.