Figure 1. CED-10/Rac1 Binds to MIG-10/Lamellipodin and Is Required for the Asymmetric Localization of MIG-10/Lamellipodin.(A) GST fusion proteins encoding activated mutants of the Rho family GTPases (RhoL63, RacL61, and Cdc42L61) were incubated with the MIG-10 RAPH domain fused to GFP (MIG-10 RAPH::GFP). MIG-10 RAPH::GFP bound to RacL61 but not Rho L63 or Cdc42L61.(B) MIG-10 RAPH::GFP was incubated with an activated Rac mutant (RacL61) or an inactive Rac mutant (RacN17). MIG-10 RAPH::GFP bound to RacL61 but not RacN17.(C) The UNC-6/netrin guidance cue is expressed from cells located ventral to the HSN and is thought to form a chemotactic gradient that polarizes the HSN [6].(D) In the wild-type background, MIG-10::YFP is localized to the ventral edge of the HSN. Previous work has indicated that this ventral localization is netrin dependant [6].(E) In the
ced-10(
n3246) mutant background, MIG-10::YFP fails to localize to the ventral edge of the HSN neuron.(F) Average ventral enrichment of MIG-10::YFP in wild-type (n = 12) and
ced-10(
n3246) (n = 15) backgrounds. Asterisk indicates p < 0.0001 (Student's t test). Myristolated::GFP (MYR::GFP) was not enriched in the wild-type or
ced-10(
n3246) background. Error bars represent standard error of the mean.Images are of collapsed stacks of optical sections. Ventral is down, and anterior is to the left. Scale bars represent 5 μm. The molecular-weight markers represent 150, 100, 75, and 50 KDa.