Fig. 8. Co-regulation of BLACAT3/YBX3 complex on NCF2 expression by binding NCF2 promoter and enhance NCF2 gene transcription, then activate NF-kB signaling pathway.

Dual-luciferase gene reporter assay (A, C) and ChIP-qPCR assay (B, D) were performed to verify that BLACAT3 can bind YBX3. E ChIP-qPCR was performed to verify BLACAT3/YBX3 complex can bind to NCF2 promoter and regulate the transcription. F, G WB detected the regulation effects of BLACAT3 and/or YBX3 knockdown on NCF2 protein level. H HALLMARK gene set enrichment analysis (GSEA) based on RNA sequencing data revealed typical signaling pathways regulated by BLACAT3. I WB verified potential signaling pathways in T24 and 5637 cells. ns no significance. Statistical significance was assessed using two-tailed Student’s t test between two groups, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.