Figure 3

A. BRMS1 downregulates OPN via the NF-κB site. A reporter plasmid bearing three NF-κB sites from the OPN promoter, pGL3-3XOPN/NF-κB, upstream of the SV40 promoter or pGL3-control was co-transfected with pCMV-mycBRMS1 into COS-7 cells. Luciferase activity from this reporter construct was measured. The experiment was repeated thrice in triplicate.* indicates significant suppression compared with respective controls (p < 0.05). B. The OPN promoter lacking an intact NF-κB site is relieved for suppression by HDAC3. The OPN promoter construct, pGL3-OPN, and its deletions, OPN-NF (retains the NF-κB-binding site) and OPN-ΔI1 (lacks the NF-κB-binding site) and the OPN/NotI construct (OPN/NF-κB site is replaced with a NotI site) were co-transfected with pcDNA3 or pcDNA3-FLAG-HDAC3 in COS-7 cells and monitored for the effect of HDAC3 on the promoter activity of OPN. The results shown represent the experiment done twice in triplicate. * indicates significant suppression compared with respective controls (p < 0.05).