Chromatin immunoprecipitation analysis was performed as described previously (Jiang et al., 2018b ). In brief, ESCC cells KYSE150, KYSE510, and TE3 were treated with THZ1 (100 nm , 12 h), and then, 1 × 107 cells were cross‐linked with 1% formaldehyde solution (Thermo Fisher Scientific) and neutralized by 1.25 m glycine. Cross‐linked cells were lysed and sonicated (Covaris E220, Woburn, MA, USA) to release 100–00 bp fragments. Anti‐KLF5 (Santa Cruz Biotechnology; sc‐398470x), anti‐TCF3 (Santa Cruz Biotechnology; sc‐166411x), or normal IgG was added to each sonicated chromatin and incubated at 4 °C overnight. Then, these complexes were conjugated to Dynabeads protein A/G magnetic beads (Invitrogen) for 4 h at 4 °C. After incubation, DNA was eluted from immunoprecipitate complexes, reverse cross‐linked, and purified with QIAquick PCR purification kit (QIAGEN, Hilden, Germany).
The purified DNA was analyzed by real‐time PCR with the use of LINC00094 super‐enhancer‐specific primers. Primers for ChIP‐PCR were shown in TableS5 . Relative enrichment was normalized to input. IgG antibody was used as a negative control.
The purified DNA was analyzed by real‐time PCR with the use of LINC00094 super‐enhancer‐specific primers. Primers for ChIP‐PCR were shown in Table