Fig. 5. Ran binds to XMog1 in co-precipitation assays. (A). Binding of XMog1 to GST-Ran. Incubations were carried out using 2 µM GST-Ran and 0.07 µM XMog1 purified proteins in Mg²⁺-buffer with addition of GTP, GDP or EDTA. A western blot of the precipitates was developed using antibodies to GST (to show the extent of recovery of GST-Ran) and XMog1. (B) Co-precipitation of XMog1 from Xenopus egg extract (XEE) by RanQ69L-GTP, RanT24N-GDP or wild-type Ran-GDP proteins (5 µM) produced as GST-fusions. GST-Ran proteins were incubated in XEE and recovered on glutathione-Sepharose beads in Mg²⁺-buffer. A western blot of the precipitates was developed using antibodies to GST (to show the extent of recovery of GST-Ran) and XMog1. (C) Co-precipitation of Ran from XEE by GST-XMog1 with the addition of GTPS or GDP. A western blot of the precipitates was developed using an antibody to Ran. Total Xenopus egg extract (XEE) was also loaded.