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Table I

Summary of the Effects of Pharmacologic and Biochemical Reagents on Dynamics of GFP–CP and GFP–Arp3

Effects
Reagent testedConcentration and durationOn spotsOn 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 
Effects
Reagent testedConcentration and durationOn spotsOn 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.  

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