TABLE 1.
F-actin quantitation in cells
| Cell line | Mean fluorescence intensitya | Total amt of F-actinb | % of total in cytoskeletonc | Fold increase |
|---|---|---|---|---|
| Ax4 parental-R3 | 488.63 | |||
| V12-RacB-R3 | 865.35 | 1.77 | ||
| Ax4 parental-R5 | 692.88 | |||
| V12-RacB-R5 | 1,082.03 | 1.56 | ||
| AX4 parental | 1 | 30.1 ± 4 | ||
| V12-RacB | 1.05 ± 0.12 | 58.9 ± 4 | 1.96 | |
| WT-RacB | 0.98 ± 0.02 | 39.9 ± 8 | 1.33 |
a
The mean intensity of fluorescence in two different size gates (R3 and R5) is shown for each cell line.
b
Total F-actin (expressed relative to the amount in parental cells) was determined by staining fixed cells with fluorescent phalloidin and then quantifying bound dye by flow cytometry.
c
Cytoskeletal actin was quantified by lysing cells, pelleting the cytoskeleton, and determining the amount of actin associated with the pellet by densitometry of Coomassie blue-stained SDS-polyacrylamide gels.