HS chains modulate SDN-1 synaptic content.(A) Black arrowheads indicate the position of putative GAG chain anchoring serines in SDN-1. Serines were mutated to alanines to generate ΔGAG mutants. Open arrowhead indicates the insertion site of AID::mNG fluorescence tag. (B) Western blot analysis (anti-mNG) of nonmutated SDN-1-mNG (lane 2), SDN-1(Δ3GAG)-mNG (lane 3), SDN-1(Δ2GAG)-mNG (lane 4), or SDN-1(Δ1GAG)-mNG (lane 5) proteins precipitated from worm lysate using mNG beads. Expected SDN-1-mNG molecular weight is 58 kD. Bracket likely indicates the SDS-resistant dimerization form of SDN-1. Arrowhead indicates potential cleaved C terminus–mNG band. Protein samples from lane 6 (shown in red) were digested by heparinase I and III. Protein lysate from N2 wild-type animals was used as control in lane 1. (C) Confocal detection and quantification of SDN-1 levels from SDN-1-mNG and ΔGAG mutants. Scale bar, 10 µm.
Figure 3.

HS chains modulate SDN-1 synaptic content. (A) Black arrowheads indicate the position of putative GAG chain anchoring serines in SDN-1. Serines were mutated to alanines to generate ΔGAG mutants. Open arrowhead indicates the insertion site of AID::mNG fluorescence tag. (B) Western blot analysis (anti-mNG) of nonmutated SDN-1-mNG (lane 2), SDN-1(Δ3GAG)-mNG (lane 3), SDN-1(Δ2GAG)-mNG (lane 4), or SDN-1(Δ1GAG)-mNG (lane 5) proteins precipitated from worm lysate using mNG beads. Expected SDN-1-mNG molecular weight is 58 kD. Bracket likely indicates the SDS-resistant dimerization form of SDN-1. Arrowhead indicates potential cleaved C terminus–mNG band. Protein samples from lane 6 (shown in red) were digested by heparinase I and III. Protein lysate from N2 wild-type animals was used as control in lane 1. (C) Confocal detection and quantification of SDN-1 levels from SDN-1-mNG and ΔGAG mutants. Scale bar, 10 µm.

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