Nuclear translocation of endogenous MRTF A after force application
| . | . | Cells exhibiting nuclear staining for endogenous MRTF A (%) . | . | |
|---|---|---|---|---|
| Time (minutes) . | Untreated . | Latrunculin B (1 μM, 30 minutes before force treatment) . | Y27632 (10 μM, 30 minutes before force treatment) . | |
| 0 | 6±2 | 5±2 | 4±2 | |
| 15 | 12±3 | 6±2 | 7±3 | |
| 30 | 27±5 | 9±3 | 10±4 | |
| 45 | 48±6 | 11±4 | 12±4 | |
| . | . | Cells exhibiting nuclear staining for endogenous MRTF A (%) . | . | |
|---|---|---|---|---|
| Time (minutes) . | Untreated . | Latrunculin B (1 μM, 30 minutes before force treatment) . | Y27632 (10 μM, 30 minutes before force treatment) . | |
| 0 | 6±2 | 5±2 | 4±2 | |
| 15 | 12±3 | 6±2 | 7±3 | |
| 30 | 27±5 | 9±3 | 10±4 | |
| 45 | 48±6 | 11±4 | 12±4 | |
Cells were incubated with collagen beads and force was applied for 0, 15, 30 or 45 minutes. Cells were fixed and stained for endogenous MRTF A using a polyclonal antibody generously donated by H. Nakano (Juntendo University School of Medicine, Tokyo, Japan). Percentage of cells that exhibited nuclear staining for endogenous MRTF A was evaluated by fluorescence microscopy. A significantly higher percentage of MRTF-A-stained nuclei was found 15, 30 or 45 minutes after force application in untreated (control cells) compared with those treated with latrunculin B or Y27632 (P<0.05)