Fig. 8.

KLF5 and TCF3 bind to SE regions for regulation of LINC00094. (A) The ranked transcription factors according to number of SE‐associated ce‐lncRNAs significantly regulated by transcription factors. (B) H3K27ac ChIP‐seq signals at the LINC00094 locus in six ESCC cell lines. Three constituent enhancers (E1, E2, and E3) within the SE were labeled in grey shadings. TCF3 and KLF5 motif occupy at E1, E2, and E3 enhancer loci. (C) ChIP‐qPCR experiments measuring TCF3 and KLF5 binding enrichment on the LINC00094 SEs segments (divided into enhancer 1, E1; enhancer 2, E2 and enhancer 3, E3) upon treating with THZ1 (100 nm,12 h). Two pairs of primers were designed for each SE segments, which has the better enrichment was finally selected. (D) Relative RNA expression of LINC00094 upon knockdown of TCF3 or KLF5 in KYSE150, KYSE150, and TE3 cells. (E) Western blotting analysis of KLF5 and TCF3 expression in KYSE150, KYSE510 and TE3 cells which were treated with either THZ1 or DMSO at indicated time points and indicated concentrations. Bars of D represent mean ± SD of three experimental replicates. *P < 0.05, **P < 0.01, ***P < 0.001. P values were determined using t‐test.