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. 2020 Jun 22;219(8):e201907184. doi: 10.1083/jcb.201907184

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© 2020 Martinez et al.

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Figure S1. — HIS-TAN1-GFP and HIS-TAN1-GFP-Atto488 binding affinity to Taxol-stabilized microtubules. (A) HIS-TAN1-GFP cosedimentation binding data with fits to hyperbolic binding isotherms for HIS-TAN1 (replotted from Fig. 1 C), HIS-TAN1-GFP, and HIS-TAN1-GFP-Atto488. Apparent affinity for HIS-TAN1-GFP is 0.595 µM ± 0.389 to 0.800 µM, while HIS-TAN1-GFP-Atto488 is 0.05 µM ± 0.0009 to 0.129 µM corrected for the average pelleting in samples without microtubules added (average ± 95% CI). (B) Coomassie-stained SDS-PAGE experiment from spindown of HIS-TAN1-GFP in the presence of varying concentrations of tubulin (0–8 µM), alternating soluble (S) and pellet (P) fractions . (C) Coomassie-stained SDS-PAGE experiment from spindown of HIS-TAN1-GFP-Atto488 in the presence of varying concentrations of tubulin (0–8 µM). Below the Coomassie-stained SDS-PAGE experiment, HIS-TAN1-GFP-Atto488 was excited using a UV light source to confirm Atto488 maleimide conjugation with HIS-TAN1-GFP used in the spindown assays.