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Figure 4


Fig. 4. INPP5B interacts with RAB proteins associated with the secretory and endocytic pathways. (A) Full-length INPP5B was tested for interaction with the indicated GTP-locked (Q to L) or GDP-locked (S or T to N) RAB proteins. Growth on high selection indicates interaction. (B) Beads containing NusA or GST alone or GST-tagged RAB proteins 1, 2, 4, 5, 6, 9, 11, 14 and 33B or NusA-tagged RAB 8 were incubated with buffer alone (`B') or HeLa cell extract containing GFP-INPP5B (`C'), and bound proteins were detected by western blotting. (C) Beads containing GST-tagged versions of the indicated RAB proteins were incubated with buffer alone (`B') or HeLa cell extract containing GFP-INPP5B and bound GFP-INPP5B detected by western blotting. (D) Beads containing GST-tagged versions of the indicated GTP-locked RAB proteins were incubated with buffer alone (`B') or recombinant full-length INPP5B (`I'), and bound protein detected by western blotting. (E) The indicated truncated forms of INPP5B were tested for interaction with RAB5Q79L in the yeast two-hybrid system. Growth on high selection indicates interaction.





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