Figure 6

Cyclic stretch increases NF-κB-binding activity and NF-κB protein phosphorylation. (A) Representative EMSA showing protein binding to the NF-κB oligonucleotide in nuclear extracts of cardiomyocytes after cyclic stretch for various periods of time and in the presence of inhibitors. Similar results were found in another two independent experiments. Cold oligo means unlabeled NF-κB oligonucleotides. A supershifted complex is observed after addition of p65 antibody. (B) Representative Western blots for phosphorylated and total NF-κB in cardiomyocytes after stretch by 20% for various periods of time and in the presence of SB203580, losartan, and TNF-α antibody. (C) Quantitative analysis of phosphorylated NF-κB protein levels. The values from stretched cardiomyocytes have been normalized to matched C23 and corresponding total protein measurement and then expressed as a ratio of normalized values to each phosphorylated protein in control cells. Data are from 4 independent experiments. *P < 0.001 vs. control. **P < 0.01 vs. control. ⁺P < 0.001 vs. stretch 2 h. ^‡ P < 0.001 vs. stretch 2 h.