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. 2011 Oct 3;195(1):71–86. doi: 10.1083/jcb.201011112

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© 2011 Willenborg et al.

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Figure 7. — FIP5 induces SNX18- and SNX9-dependent liposome tubulation. (A) SNX18 or FIP5 were incubated with PS/PC liposomes containing various phosphatidylinositides. Liposomes were then sedimented and levels of bound SNX18 or FIP5 were determined by Coomassie staining. (B) C- (FIP5 C-terminal) or N-terminal (FIP5-C2) domains of FIP5 were tested for their ability to bind PS/PC liposomes containing 5% PI(4,5)P₂. The levels of bound proteins were determined by Coomassie staining. (C) SNX18 was incubated with PS/PC/PI(4,5)P₂ liposomes in the presence or absence of GST-FIP5. The levels of bound SNX18 and GST-FIP5 were determined by Coomassie staining. Black lines indicate the removal of intervening lanes for presentation purposes. (D) EM analysis of liposomes incubated with GST, GST-SNX9, GST-SNX18, 6His-FIP3, and 6His-FIP5. Arrows point to wide tubules (122.1 ± 32.9 nm, n = 10) and arrowheads point to narrow tubules (54.1 ± 18.1nm, n = 10). The asterisk marks short tubules induced by SNX18 alone.