Fig. 4

Verification of the regulatory roles of rs10896081. a Disruption of PBX3 binding by the SNP rs10896081. b ChIP-Seq tracks showing DNase-Seq signals (light blue), TF ChIP-Seq signals (green), and histone modifications (purple) near rs10896081. c Reporter gene assays showed that the T allele of rs10896081 produced significantly higher luciferase activity than the A allele in all three tested cell lines. d–f Knockdown of PBX3 increased the expression of PACS1 and decreased the expression of YIF1A, indicating that PACS1 and YIF1A are regulated by PBX3. g The SNP rs10896081 is located in the first intron of the longest transcript of PACS1. h–j Deletion of the genomic region containing rs10896081 led to dysregulation of PACS1 and YIF1A. h Electrophoretic analysis showed that the segment containing rs10896081 was deleted from the genome. The expected DNA length of rs10896081 in wild-type cells (WT) was 1042 bp, and that in edited cells (KO) was 646 bp. UTR, untranslated region. CDS, coding sequence. N = 8 per group for HEK293T cells, n = 8 for the control group, n = 16 per experimental group for SH-SY5Y and U251 cells, n = 3 per group in (d–f) and (h–k). Two-tailed Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001