Figure 2.
R8-9 and R4 or R10 displayed opposite preferences for tubulin dimer spacing. (A) Images showing the preferential binding of GFP-LZ-TPX2 R8-9 (5 nM) to curved segments of dynamic GDP-MT lattices and MT end in a flow-in assay. (B) Line-scan intensity profiles of TPX2 R8-9 (green) and MT (red) corresponding to the dashed curved lines in A. (C) Quantification of intensities of 5 nM GFP-LZ-TPX2 R8-9 on straight GDP-MT, curved GDP-MT, MT ends, and GMPCPP-MT. n = 47–50 MTs from two experiments. (D and E) TIRF microscopy images of live MRC5 cells expressing GFP-LZ-TPX2 R8-9 together with mCherry-α-tubulin before (upper panels) or after (lower panels) the addition of 1 µM paclitaxel (D). Quantification of the fluorescence intensities of GFP-LZ-TPX2 R8-9 on MTs without or with paclitaxel treatment (E). n = 4 experiments (∼70 MTs were measured). ***P < 0.001, two-tailed t test (paired). (F–H) Images showing the behavior of GFP-LZ–tagged TPX2 R8-9 (F), R4 (G), and R10 (H) (green) at 200 nM or 2 µM on dynamic MTs (red) in the absence or the presence of 1 µM SiR-tubulin (blue). (I) Quantification of intensities of GFP-LZ–tagged TPX2 R8-9, R4, and R10 at 200 nM or 2 µM on GMPCPP seeds and GDP-MT lattices. n = 20 MTs from two experiments. N.D., not detected. (J) Images showing the behavior of GFP-LZ–tagged TPX2 R8-9 (green) at 200 nM on dynamic MTs (blue) alone or together with mCherry-LZ–tagged TPX2 R4 (red) at 200 nM or 1 µM. (K) Quantification of the intensities of GFP-LZ–tagged TPX2 R8-9 on GMPCPP seeds and GDP-MT lattices. n = 20 MTs from two experiments. Scale bars, 2 μm. Data represent mean ± SD. Refer to the image caption for details.

R8-9 and R4 or R10 displayed opposite preferences for tubulin dimer spacing. (A) Images showing the preferential binding of GFP-LZ-TPX2 R8-9 (5 nM) to curved segments of dynamic GDP-MT lattices and MT end in a flow-in assay. (B) Line-scan intensity profiles of TPX2 R8-9 (green) and MT (red) corresponding to the dashed curved lines in A. (C) Quantification of intensities of 5 nM GFP-LZ-TPX2 R8-9 on straight GDP-MT, curved GDP-MT, MT ends, and GMPCPP-MT. n = 47–50 MTs from two experiments. (D and E) TIRF microscopy images of live MRC5 cells expressing GFP-LZ-TPX2 R8-9 together with mCherry-α-tubulin before (upper panels) or after (lower panels) the addition of 1 µM paclitaxel (D). Quantification of the fluorescence intensities of GFP-LZ-TPX2 R8-9 on MTs without or with paclitaxel treatment (E). n = 4 experiments (∼70 MTs were measured). ***P < 0.001, two-tailed t test (paired). (F–H) Images showing the behavior of GFP-LZ–tagged TPX2 R8-9 (F), R4 (G), and R10 (H) (green) at 200 nM or 2 µM on dynamic MTs (red) in the absence or the presence of 1 µM SiR-tubulin (blue). (I) Quantification of intensities of GFP-LZ–tagged TPX2 R8-9, R4, and R10 at 200 nM or 2 µM on GMPCPP seeds and GDP-MT lattices. n = 20 MTs from two experiments. N.D., not detected. (J) Images showing the behavior of GFP-LZ–tagged TPX2 R8-9 (green) at 200 nM on dynamic MTs (blue) alone or together with mCherry-LZ–tagged TPX2 R4 (red) at 200 nM or 1 µM. (K) Quantification of the intensities of GFP-LZ–tagged TPX2 R8-9 on GMPCPP seeds and GDP-MT lattices. n = 20 MTs from two experiments. Scale bars, 2 μm. Data represent mean ± SD.

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