Table I.
Summary of the Effects of Pharmacologic and Biochemical Reagents on Dynamics of GFP–CP and GFP–Arp3
| Effects | ||||||
|---|---|---|---|---|---|---|
| Reagent tested | Concentration and duration | On spots | On the cell periphery | |||
| Inhibitors of actin polymerization | ||||||
| Cytochalasin D | 1 μM, 2 min | Motility inhibited | Motility inhibited | |||
| Latrunculin B | 200 nM, 2 min | Motility inhibited | Motility inhibited | |||
| Swinholide A | 50 nM, 5 min | Motility inhibited | Motility inhibited | |||
| Jasplakinolide | 1.4 μM, 15 min | Motility inhibited | Motility inhibited | |||
| Microtubule reagents | ||||||
| Nocodazole | 1 μM, 30 min | No effect | No effect | |||
| Taxol | 10 nM, overnight | No effect | No effect | |||
| Myosin inhibitors | ||||||
| Butanedione monoxime | 10 mM, 15–60 min | No effect | n.d.* | |||
| KT5926 | 100 nM, 15–60 min | No effect | No effect | |||
| Kinase and phosphatase inhibitors | ||||||
| Okadaic acid | 100 nM, 30 min | No effect | No effect | |||
| Genistein | 25 μM, 30 min | No effect | No effect | |||
| Expression/modification of small G proteins | ||||||
| N17Rac | 2 h after cDNA injected | CP and Arp3 diffuse | Motility inhibited | |||
| V12Rac | 2 h after cDNA injected | No effect | No effect | |||
| N17cdc42 | 2 h after cDNA injected | CP and Arp3 diffuse | Motility inhibited | |||
| V12cdc42 | 2 h after cDNA injected | No effect | No effect | |||
| V14RhoA | 15–60 min after injection at 300 μg/ml | No effect | No effect | |||
| Exoenzyme C3 transferase | 10–50 min after injection at 200 μg/ml | CP and Arp3 diffuse | Motility inhibited | |||
| Growth factors and mitogens | ||||||
| Phorbol myristic acid | 2 μM, 30 min | No effect | Motility increased | |||
| EGF | 500 ng/ml, 60 min | No effect | No effect | |||
| Reagents that alter phosphoinositide metabolism | ||||||
| Wortmannin | 100 nM, 60 min | No effect | No effect | |||
| Anti-PIP2 | 15–60 min after injection at 11 mg/ml | No effect | No effect | |||
| PIP2-binding peptide | 15–60 min after injection at 10 mM | No effect | No effect | |||
| PI 5-kinase | 2 h after cDNA injected | Increased movement | n.d. | |||
| Effects of cofilin regulators | ||||||
| LIMK-1 | 2 h after cDNA injected | Inhibited movement | Inhibited | |||
| Mutant LIMK-1 | 2 h after cDNA injected | No effect | No effect | |||
*
n.d., not determined. Changes in morphology at the cell periphery made it difficult to observe motility at the edge of the cell.