(A) Venn diagram presenting potential transcription factors binding to the SE regions (E1 and E2) and the core promoter region of EFNA1. (B) Pearson’s correlation analysis showing the positive correlation between EFNA1 and FOSL2 expression in CC samples from the TCGA database (27). TPM, transcripts per million. (C) Expression of FOSL2 and EFNA1 in SiHa and HCC-94 cells transiently transfected with siRNAs targeting FOSL2 or control siRNA. mRNA expression was determined using qRT-PCR. (D and E) Western blotting showing the protein expression levels of FOSL2 and EFNA1 following FOSL2 knockdown (D) or overexpression (E). (F and G) Luciferase reporter assay showing EFNA1 promoter activity in HEK293T cells with FOSL2 knockdown (F) or overexpression (G). (H) Gene tracks showing FOSL2 ChIP-Seq occupancy at the EFNA1 loci in SiHa and HCC-94 cell lines. The x axis shows genomic position, and the y axis shows ChIP-Seq occupancy signal in reads per million mapped reads per base pair (rpm/bp). (I–K) qPCR assay showing FOSL2 and H3K27ac enrichments at the EFNA1 core promoter and SE regions from ChIP assay in HCC-94 cells with FOSL2 and H3K27ac antibodies. Data are presented as mean ± SD from 3 independent experiments. (L) Schematic diagram showing the EFNA1 E2 region (from chr1:155,100,545–155,103,815) with an FOSL2 binding motif (from chr1:155,101,975–155,101,985). (M) Luciferase activity of the indicated plasmids in HEK293T cells. After 48 hours of transfection of specified plasmid, luciferase activity was determined and normalized to pRL-TK luciferase activity. Data are presented as mean ± SD across n = 3 replicates. Statistical analysis was performed using Pearson’s correlation test in B, 1-way ANOVA test in C, F, G, I–K, and M. Significant P values: **P < 0.01, ***P < 0.001.