Table I.
Frequencies and Extents of MTOC- and Periphery-directed ES and No Movement Frequencies (0) of Wt and Ts1 Ad2 in TC7, TC7/MAP4/MTB–GFP, and HeLa Cells
| MTOC | Periphery | 0 | Total | |||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Peak | Mean | ES | Peak | Mean | ES | ES | Mean | ES | Particles | Cells | ||||||||||||
| μm/s | μm/s | μm/s | μm/s | μm/s | ||||||||||||||||||
| TC7 Contr | ||||||||||||||||||||||
| No drug | 2.06 | 0.50 ± 0.34 | 295 (40.5%) | 2.33 | 0.42 ± 0.23 | 227 (31.1%) | 207 (28.4%) | −0.06 ± 0.22 | 729 | 15 | 8 | |||||||||||
| TC7/MAP4-G | ||||||||||||||||||||||
| No drug | 2.63 | 0.50 ± 0.15 | 531 (55.4%) | 1.38 | 0.31 ± 0.11 | 304 (31.7%) | 123 (12.8%) | −0.21 ± 0.19 | 958 | 19 | 6 | |||||||||||
| Tax | 2.12 | 0.44 ± 0.19 | 340 (35.4%) | 1.08 | 0.33 ± 0.23 | 228 (23.8%) | 392 (40.8%) | −0.14 ± 0.15 | 960 | 18 | 7 | |||||||||||
| Surface | 0.19 | 0.12 ± 0.02 | 76 (8.3%) | 0.20 | 0.11 ± 0.01 | 71 (7.8%) | 769 (84.0%) | −0.00 ± 0.00 | 916 | 12 | 5 | |||||||||||
| HeLa | ||||||||||||||||||||||
| No drug | 2.08 | 0.31 ± 0.14 | 555 (33.1%) | 2.19 | 0.33 ± 0.15 | 427 (25.5%) | 695 (41.4%) | −0.03 ± 0.17 | 1,677 | 28 | 11 | |||||||||||
| DynGFP | 2.13 | 0.22 ± 0.10 | 143 (17.7%) | 1.08 | 0.30 ± 0.12 | 255 (31.5%) | 412 (50.9%) | 0.06 ± 0.10 | 810 | 25 | 9 | |||||||||||
| GFP | 1.36 | 0.36 ± 0.11 | 330 (37.3%) | 1.45 | 0.34 ± 0.19 | 228 (25.8%) | 327 (36.9%) | −0.05 ± 0.24 | 885 | 14 | 13 | |||||||||||
| Noc | 0.28 | 0.15 ± 0.02 | 65 (10.7%) | 0.36 | 0.17 ± 0.05 | 82 (13.5%) | 461 (75.8%) | 0.04 ± 0.09 | 608 | 13 | 3 | |||||||||||
| Surface | 0.17 | 0.11 ± 0.01 | 24 (5.4%) | 0.17 | 0.11 ± 0.02 | 24 (5.4%) | 400 (89.3%) | 0.00 ± 0.01 | 448 | 16 | 4 | |||||||||||
| Ts1 Ad | ||||||||||||||||||||||
| TC7 Contr | ||||||||||||||||||||||
| No drug | 2.48 | 0.36 ± 0.10 | 544 (35.5%) | 2.12 | 0.39 ± 0.12 | 641 (41.9%) | 346 (22.6%) | 0.04 ± 0.08 | 1,531 | 15 | 7 | |||||||||||
| TC7/MAP4-G | ||||||||||||||||||||||
| No drug | 2.28 | 0.33 ± 0.14 | 573 (40.6%) | 1.84 | 0.30 ± 0.14 | 520 (36.9%) | 317 (22.5%) | −0.02 ± 0.14 | 1,410 | 32 | 5 | |||||||||||
| Tax | 2.69 | 0.45 ± 0.19 | 422 (38.8%) | 1.84 | 0.39 ± 0.23 | 369 (33.9%) | 298 (27.4%) | −0.07 ± 0.18 | 1,089 | 19 | 3 | |||||||||||
Motilities of TR-labeled virus particles in control or drug-treated cells were recorded by time-lapse fluorescence microscopy using intervals between 1.2 and 2.6 s and analyzed as described in Materials and Methods. ES were calculated from the elementary motion steps of selected virus particles that were located well within the cell and were able to move in at least five subsequent frames over a minimal distance of 0.5 μm. ES were grouped according to their directionalities, MTOC (minus end), periphery (plus end), and no motility (0), and characterized in terms of peak speeds, mean speeds, and frequencies. The no motility category comprised movements of less than 0.1 μm/s, which was within the systematic error of our measurements. The total amounts of ES was between 448 and 1,677 and derived from three to seven independent experiments analyzing 13–32 different virus particles in 3–13 different cells, respectively. The mean plus- and minus end–directed speeds including the standard deviations were determined by averaging the corresponding mean velocities of individual particles. Total mean speeds (representing the population speeds) including the standard deviations were calculated for each particle that was traced. Minus values represent MTOC-directed speeds, positive values are periphery-directed speeds. (For significant differences between different mean speeds, see Table II.) Movement frequencies were different in all cases listed above, except in the following instances (as analyzed in a chi square test). With wt Ad2: HeLa control versus HeLa GFP (peripheral, MTOC, and no motility); TC7 control versus TC7/MAP4/MTB–GFP (peripheral speeds); TC7/MAP4/MTB–GFP taxol versus HeLa control or HeLa GFP (peripheral and MTOC speeds). With ts1 Ad2: TC7/MAP4/MTB–GFP control versus TC7/MAP4/MTB–GFP taxol (periphery, MTOC).