Figure 2. EB1 is necessary and... | Rockefeller University Press

Figure 2.

$Figure 2. EB1 is necessary and sufficient for end tracking of CLIP-170 in buffer. (A–C) TIRF microscopy images (top) and kymographs (bottom) of dynamic Alexa Fluor 568–labeled microtubules (red) in buffer in the presence of the following purified GFP-labeled +TIPs (green): 50 nM CLIP-170–GFP (A, left) or 50 nM H2-GFP (A, right); 150 nM EB1-GFP (B); and 35 nM CLIP-170–GFP (C, left) or 35 nM H2-GFP (C, right) in the presence of 70 nM of unlabeled EB1 in the presence of 15.5 μM tubulin. Kymographs display a period of 5 min. Bars, 5 μm. (D and E) Analytical gel filtrations: UV absorbance profiles and the corresponding Coomassie-stained SDS gel fractions of runs of a mixture of 20 nM H2 and 40 nM GTP-tubulin (D, red), and of 20 nM H2, 20 nM EB1, and 40 nM GTP-tubulin (E, red). Runs of H2 alone (blue), GTP-tubulin alone (green), and EB1 alone (black) at concentrations as in the mixtures are shown for comparison.$

EB1 is necessary and sufficient for end tracking of CLIP-170 in buffer. (A–C) TIRF microscopy images (top) and kymographs (bottom) of dynamic Alexa Fluor 568–labeled microtubules (red) in buffer in the presence of the following purified GFP-labeled +TIPs (green): 50 nM CLIP-170–GFP (A, left) or 50 nM H2-GFP (A, right); 150 nM EB1-GFP (B); and 35 nM CLIP-170–GFP (C, left) or 35 nM H2-GFP (C, right) in the presence of 70 nM of unlabeled EB1 in the presence of 15.5 μM tubulin. Kymographs display a period of 5 min. Bars, 5 μm. (D and E) Analytical gel filtrations: UV absorbance profiles and the corresponding Coomassie-stained SDS gel fractions of runs of a mixture of 20 nM H2 and 40 nM GTP-tubulin (D, red), and of 20 nM H2, 20 nM EB1, and 40 nM GTP-tubulin (E, red). Runs of H2 alone (blue), GTP-tubulin alone (green), and EB1 alone (black) at concentrations as in the mixtures are shown for comparison.