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Data Supplement for Circulation Research: Volume 86, Issue 5 -- Page 549
This item has the following additional materials available:
Data Files
- Figure 1. Time-lapse fluorescence microscopy of MAEC at the wound margin, loaded with CM-DCF-DA 1 hr after wounding. The wound is toward the left side. Images were obtained using the 40x objective, and the MetaMorph software, on a SenSys digital camera, with no binning. The parameters were as follows: images were recorded every 90 sec, at 490 nm excitation wavelength, 1 sec exposure, with the ND8 and ND16 filters in the light pass. Note the increasing fluorescence in the actively ruffling bottom cell. Also note that the ruffling activity is not synchronized in all cells, as the top cell is more quiescent.
- Figure 7. Time-lapse microscopy of a wounded monolayer of MAEC. The cells were maintained in DMEM supplemented with 10% fetal bovine serum, to which a 1:1000 dilution of vehicle (DMSO) was added 1 hr prior to wounding and for the entire duration of the experiment. Images were recorded every 90 sec for 5 hr, using the 40x objective and the MetaMorph software. Note the intense ruffling and pinocytotic activity of most of the migrating cells at the wound margin.
- Figure 7. Time-lapse microscopy of a wounded monolayer of MAEC, pretreated for 1 hr prior to wounding with the NADPH oxidase inhibitor DPI (10 µmol/L), in the same culture medium as the control. The inhibitor was maintained in the medium for the entire duration of the experiment. Time-lapse was performed as in control. Note the reduced speed and the more random migration of the cells, as well as the lack of pinocytotic activity. The ruffling activity is also much reduced, and the lamellipodia have a different appearance.
- Figure 7. Time-lapse microscopy of a wounded monolayer of MAEC, pretreated for 1 hr prior to wounding with the superoxide dismutase mimetic MnTMPyP (25 µmol/L), in the same culture medium as control cells. The scavenger was maintained in the medium for the entire duration of the experiment. Time-lapse was performed as in control. The speed and directionality of the cellular movement are less than in control, while the pinocytotic activity is absent. The ruffling activity is also reduced, and the lamellipodia have the same appearance as in DPI-treated cells.
Prepared by: the Data Supplement Manager
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