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. 2015 Nov 30;13(1):98–108. doi: 10.1080/15476286.2015.1122164

Figure 4.

Figure 4.

ANRIL binds with YY1 to cooperatively activate IL6 and IL8 expression upon TNF-α treatment. (A) YY1 binding sites in promoter regions of human IL6 and IL8 genes according to UCSC Genome Browser (http://genome.ucsc.edu). TSS means transcriptional start site. (B and C) YY1 (B) and IL6, IL8 (C) levels in HUVECs transfected with scramble or 2 sets of YY1 siRNAs (siYY1–1 and siYY1–2, 24h) upon TNF-α treatment (25 ng/mL, 24h). (*p < 0.05, **p < 0.01, ***p < 0.005). (D) RIP analysis showing YY1 binding with ANRIL upon TNF-α treatment (25 ng/mL, 6h). GAPDH RNA was used as a control. (*p < 0.05, n.s. means non-significant). (E) CHIP assay showing that ANRIL knockdown by siRNA transfection (24h) impaired YY1 binding to IL6, IL8 and CXCR4 promoter regions upon TNF-α treatment (25 ng/mL, 6h). GAPDH DNA was used as a control. (*p < 0.05, **p < 0.01, n.s. means non-significant). (F) CHIRP assay showing that ANRIL bound to IL6, IL8 and CXCR4 promoter regions upon TNF-α treatment (25 ng/mL, 6h). ANRIL probes were used and NC probe was used as negative probe control. TERC DNA was used as a control. (*p < 0.05, n.s. means non-significant). All results are presented as mean ± SEM from at least 3 independent biological experiments.