Movie 7

Cofilin inhibitory peptides prevent MTOC polarization towards anti-Ig-coated coverslips. A20 cells expressing mTagRFP-tubulin (red) and GFP-F-tractin (green) were treated with the control Q peptide (5 μM) or the M/W cofilin-inhibitory peptides (5 μM each) and added to anti-IgG-coated coverslips. Starting at 5 min after adding the cells to the coverslips, time-lapse spinning disk confocal microscopy was used to acquire z-stacks at 0.5 μm step size once every 22 s for a total of 7 min for Q peptide-treated cells, and at 0.75 μm step size every 45s for a total of 18 min for M/W peptide-treated cells. 3D reconstruction of z-stacks with side and top views of the cells are played back at 4 frames per second (~100X real time for Q peptide-treated cells and 180X real time for M/W peptide-treated cells).

The Rap1–cofilin-1 pathway coordinates actin reorganization and MTOC polarization at the B cell immune synapse

Jia C. Wang, Jeff Y.-J. Lee, Sonja Christian, May Dang-Lawson, Caitlin Pritchard, Spencer A. Freeman, and Michael R. Gold

J Cell Sci 2017. 130:1094-1109; doi: 10.1242/jcs.191858