Fig. 5

CTCF-s activates IFI6 by disrupting CTCF-mediated looping and establishing distal enhancer−promoter contacts. a GO analysis of upregulated genes after CTCF-s gain-of-function. b Relative expression of interferon-signaling-associated genes after CTCF and CTCF-s gain-of-function, respectively (mean ± s.d., two-tailed Student’s t test; n = 3 biological replicates, each biological replicate had three technical replicates). c Growth curves of HeLa-S3 cells with gain-of-function of FLAG, FLAG-CTCF and FLAG-CTCF-s, respectively. Data were shown as mean ± s.d. from three independent experiments with the indicated significance by using a two-tailed Student’s t test. d Effects of CTCF and CTCF-s overexpression on apoptosis in HeLa-S3 cells after knocking-down endogenous CTCF and CTCF-s. Data were shown as mean ± s.d., n = 3 biological replicates. e Apoptosis analysis of HeLa-S3 cells overexpressing FLAG, FLAG-CTCF and FLAG-CTCF-s following IFI6 knocking-down and treatment with oxaliplatin for 24 h. Data were shown as mean ± s.d. from two independent biological replicates. f Hi-C interaction map at IFI6 locus. g Genomic view showing CTCF ChIP-seq and RNA-seq data from FLAG and FLAG-CTCF-s, H3K4me3, H3K27ac and DNase-seq data at the IFI6 locus. h Verification of CTCF ChIP-seq data in IFI6 locus by using ChIP-qPCR. IgG was used as a negative control. The primers (from P6 to P9) were indicated in (g). The data were represented as mean ± s.d. with the indicated significance from a two-tailed Student’s t test, n = 3 technological replicates. i CTCF HiChIP interactions at IFI6 locus displayed with CTCF ChIP-seq and one-dimensional track of CTCF HiChIP in FLAG and FLAG-CTCF-s overexpressing cells. The directionality of the motifs at CTCF loop anchors was indicated with red arrows and gray bars. j 3C assay measuring the crosslinking frequency in FLAG and FLAG-CTCF-s overexpressing HeLa-S3 cells. IFI6 promoter (anchor) and six complementary primers (C1 to C6 in (g)) were tested by 3C-qPCR. The data were represented as mean ± s.d. with the indicated significance by using a two-tailed Student’s t test, n = 3 technological replicates. For statistical significance, *p < 0.05, **p < 0.01, ***p < 0.001, n.s. not significant. Source data are provided as a Source Data file