![Recombinantly expressed TAN1 binds to microtubules (MTs).(A) Coomassie-stained SDS PAGE results from microtubule cosedimentation with HIS-TAN1, positive control (microtubule-associated protein fraction [MAPF], 70% MAP2) and negative control (BSA) controls separated into corresponding pellet and soluble fractions. (B) Coomassie-stained SDS-PAGE results from microtubule cosedimentation assay using 2 µM HIS-TAN1 and from 0 to 8 µM tubulin, alternating soluble and pellet fractions. (C) Hyperbolic fit of microtubule cosedimentation data for HIS-TAN1 at varying concentrations of microtubules determines an apparent K0.5 value of 1.08 µM (95% CI, 0.722-1.43 µM), removing the outlier at 4 µM tubulin; error bars are 95% confidence intervals. (D and E) Rhodamine-labeled, Taxol-stabilized microtubule in buffer only control (magenta; D) or incubated with HIS-TAN1-GFP-Atto488 (green). (F and G) Time lapse and kymograph of rhodamine-labeled GMPCPP-stabilized microtubules (magenta) and HIS-TAN1-GFP-Atto488 (green). Scale bars, 10 µm.](https://cdn.rupress.org/rup/content_public/journal/jcb/219/8/10.1083_jcb.201907184/4/jcb_201907184_fig1.png?Expires=1772836830&Signature=SeVE~t~voWIMy3M9zugrq1qpJyb1ybMCf7VzkWpFG10Z2Oak28FvHv0gz94GTzKbttgFzlpWnqUwTMzKKNhFAgMoOrwEQAInf~TLT~bg~veklEj~c5huLk1cLHy~KSH3Q0qS5~OV~BVhUioVWsaYE9QrIyT-Ff9wolO1BeGXfYry3OuExE7w8fdF8ZStgoCf41eakMoi7YKGXN8v5v2OFc-aSaVPz-s4KwMwORqBl9UR2T7gkSFZ-BA6h-fe4Rn2b-RkWEtHDIq62ROz~FznlNQMyE~CTLpnUA-Ew9M75Ptg5I2qg9q951~DJcbAnqh41PANQakcjj97Np~RsMo3Bg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Recombinantly expressed TAN1 binds to microtubules (MTs). (A) Coomassie-stained SDS PAGE results from microtubule cosedimentation with HIS-TAN1, positive control (microtubule-associated protein fraction [MAPF], 70% MAP2) and negative control (BSA) controls separated into corresponding pellet and soluble fractions. (B) Coomassie-stained SDS-PAGE results from microtubule cosedimentation assay using 2 µM HIS-TAN1 and from 0 to 8 µM tubulin, alternating soluble and pellet fractions. (C) Hyperbolic fit of microtubule cosedimentation data for HIS-TAN1 at varying concentrations of microtubules determines an apparent K0.5 value of 1.08 µM (95% CI, 0.722-1.43 µM), removing the outlier at 4 µM tubulin; error bars are 95% confidence intervals. (D and E) Rhodamine-labeled, Taxol-stabilized microtubule in buffer only control (magenta; D) or incubated with HIS-TAN1-GFP-Atto488 (green). (F and G) Time lapse and kymograph of rhodamine-labeled GMPCPP-stabilized microtubules (magenta) and HIS-TAN1-GFP-Atto488 (green). Scale bars, 10 µm.
