ZFIN Image: Kurtzman et al., 2001, Fig. 3

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Figure Caption

Fig. 3 Zebrafish Vsx-1 is not a substrate for SUMO-1 conjugation in vitro or in COS-7 cells. Open and closed arrows indicate unconjugated MT-zVsx-1 and RanGAP1, respectively. Asterisk shows RanGAP1 conjugated to endogenous SUMO-1. (A) Xenopus RanGAP1 (lanes 1-4) or MT-zVsx-1 (lanes 6-9) were in vitro translated and labeled with [³⁵S]Met/[³⁵S]Cys. Recombinant GST-SUMO-1 and Ubc9 were added as indicated. Ubc9 (3x) was added to lanes 4, 5, 9, and 10. Western blots using anti-SUMO-1 or anti-hsUbc9 antibodies show relative levels of GST-SUMO-1 and Ubc9. (B) COS-7 cells were transfected with expression plasmids encoding RanGAP1, MT-zVsx-1, and GFP-SUMO-1. Cell lysates were immunoblotted with anti-RanGAP1 antibody (lanes 1 and 2 in upper gel), 9e10 antibody (lanes 3-5 in upper gel), or anti-GFP antibody (lower gel). Identical results were observed when GFP-SUMO-2 was substituted for GFP-SUMO-1.

Acknowledgments

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