Fig. 6

LINC00922 regulated ETS1 expression via H3K27cr. (A) qRT-PCR analysis of association between LINC00922 and ETS1 expression in CRC tissues (n = 28). (B-G) qRT-PCR analysis (B-D) and immunoblotting analysis (E-G) of ETS1 expression in HCT116 cells with LINC00922 stable knockdown (B, E), or CRC cells transfected with si-LINC00922 (C, F) or LINC00922 plasmid (D, G) for 48 h. (H) Immunofluorescence images of ETS1 (red) in HCT116 cells with LINC00922 stable knockdown. Scale bar, 100 μm. (I) Immunoblotting analysis of ETS1, H3K27cr, H3, and GAPDH levels in HCT116 cells treated with 10 mM NaCr for 48 h. (J-K) qRT-PCR (J) and immunoblotting (K) analysis of ETS1 expression in LINC00922 stable knockdown HCT116 cells supplemented with 10 mM NaCr for 48 h. (L) Image showing H3K27cr enrichment on the EST1 promoter region in HCT116 with LINC00922 transient overexpression for 48 h. (M) ChIP-qPCR assay analysis of H3K27cr enrichment on the EST1 promoter in HCT116 cells treated with LINC00922 stable knockdown or transient overexpression for 48 h. The numbers below western bands indicate the ratio of protein versus GAPDH or H3 in experimental groups to that in control group, analyzed using Image J2. Data are represented as means ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, unpaired, two-tailed, Student’s t-test