Figure 4.

Promoter binding of p65 transactivates mir-17-92 gene in H69 cells following LPS stimulation. (A) LPS induced the promoter element binding of p65 to C13orf25 gene. The schematic diagram shows three potential binding sites in the putative promoter element of to C13orf25 gene. ChIP analysis revealed increased binding of p65 to the binding site at −827, but not at −1442 and −1698, of C13orf25 promoter element in cells following LPS stimulation. Representative ChIP gels are shown in the upper panel and densitometry analysis in the lower panel. (B) H69 cells were transfected with various luciferase reporter constructs spanning the potential p65 binding sites of the C13orf25 promoter. The transfected cells were exposed to LPS in the presence or absence of SC-514. Luciferase activity was measured and presented as the ratio of the activity of the test construct with the control luciferase reporter construct. Reporter constructs containing different nucleotide genomic regions with the putative p65 binding sites were also utilized for the analysis as indicated. H69 cells were co-transfected with the pCMV-p65 to overexpress p65 and the luciferase reporter constructs for 24 h followed by measurement of luciferase activity. *P < 0.05 t-test versus the non-stimulated cells or empty pCMV vector control; ^#P < 0.05 t-test versus LPS-stimulated cells.