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. 2021 Dec 29;82(4):556–570. doi: 10.1158/0008-5472.CAN-21-1446

Figure 4.

Figure 4. The Uc003xsl.1/NKRF complex promotes NFκB target gene (IL8) transcription in TNBC cells. A and B, Heatmaps (A) and peaks (B) of NKRF genomic binding at the target sites in MDA-MB-231-P3 cells after the transduction of control or Uc003xsl.1 siRNA. A 2-kbp interval centered on the NKRF peak was shown. C, GSEA showed the enrichment of ChIP-seq promoter peaks with a significant increase in NKRF binding for the downregulated genes in MDA-MB-231-P3 cells after transfection of Uc003xsl.1 siRNA compared with si-NC. FDR, false discovery rate; FWER, family-wise error rate; NES, normalized enrichment score. D, Heatmap analysis displayed NFκB-regulated cytokine profiles in supernatants secreted by MDA-MB-231-P3 and Uc003xsl.1-depleted MDA-MB-231-P3 by Luminex multiplex cytokine assays. E, Venn diagram revealed intersection genes of the ChIP-seq, RNA-seq, and NFκB-regulated cytokine profiles in MDA-MB-231-P3 cells after transfection of Uc003xsl.1 siRNA and control siRNA. DEG, differentially expressed genes. F–H, IL8 mRNA levels (F), concentration in supernatants (G), and transcriptional activity (H) were respectively assessed by qRT-PCR, ELISA, and luciferase assay in MDA-MB-231-P3 cells after transfection of control siRNA, Uc003xsl.1 siRNA, NKRF siRNA, or both Uc003xsl.1 siRNA and NKRF siRNA. Error bars, SDs of three independent experiments. **, P < 0.01; ***, P < 0.001.

The Uc003xsl.1/NKRF complex promotes NFκB target gene (IL8) transcription in TNBC cells. A and B, Heatmaps (A) and peaks (B) of NKRF genomic binding at the target sites in MDA-MB-231-P3 cells after the transduction of control or Uc003xsl.1 siRNA. A 2-kbp interval centered on the NKRF peak was shown. C, GSEA showed the enrichment of ChIP-seq promoter peaks with a significant increase in NKRF binding for the downregulated genes in MDA-MB-231-P3 cells after transfection of Uc003xsl.1 siRNA compared with si-NC. FDR, false discovery rate; FWER, family-wise error rate; NES, normalized enrichment score. D, Heatmap analysis displayed NFκB-regulated cytokine profiles in supernatants secreted by MDA-MB-231-P3 and Uc003xsl.1-depleted MDA-MB-231-P3 by Luminex multiplex cytokine assays. E, Venn diagram revealed intersection genes of the ChIP-seq, RNA-seq, and NFκB-regulated cytokine profiles in MDA-MB-231-P3 cells after transfection of Uc003xsl.1 siRNA and control siRNA. DEG, differentially expressed genes. F–H, IL8 mRNA levels (F), concentration in supernatants (G), and transcriptional activity (H) were respectively assessed by qRT-PCR, ELISA, and luciferase assay in MDA-MB-231-P3 cells after transfection of control siRNA, Uc003xsl.1 siRNA, NKRF siRNA, or both Uc003xsl.1 siRNA and NKRF siRNA. Error bars, SDs of three independent experiments. **, P < 0.01; ***, P < 0.001.