FIGURE 7.

IL6‐AS1 promotes the modification of the histones H3K4me3 and H3K27ac on the interleukin (IL) 6 promoter, probably through EBF1. (A) Schematic representation of the putative modification markers, H3K27ac, H3K4me1, and H3K4me3 upstream of IL‐6 from the ENCODE database (https://genome.ucsc.edu/ENCODE/). (B–D) ChIP‐PCR analysis of H3K27ac (B), H3K4me1 (C), and H3K4me3 (D) on the IL‐6 promoter in HBF cells (two‐way ANOVA, n = 3 biological replicates). (E–H) HBF cells were transfected with SSIL6‐AS1 and assessed for H3K27ac (E), H3K4me3 (F), H3K4me1 (G) and RNA polymerase II (H) on the IL‐6 promoter by ChIP‐qPCR analysis (two‐way ANOVA, n = 3 biological replicates). (I–L) HBF cells were transfected with siEBF1‐1 and assessed for H3K27ac (I), H3K4me1 (J), H3K4me3 (K), and RNA polymerase II (L) on the IL‐6 promoter by chromatin immunoprecipitation‐qPCR analysis (two‐way ANOVA, n = 3 biological replicates). (M and N) HBF cells were cotransfected with siEBF1‐1 and IL6‐AS1 overexpression vector and assessed for H3K27ac (M) and H3K4me3 (N) on the IL‐6 promoter by ChIP‐qPCR analysis (two‐way ANOVA, n = 3 biological replicates). Error bars represent means ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001