Figure 2. KIF14 recruits Radil on... | Rockefeller University Press

Figure 2.

$Figure 2. KIF14 recruits Radil on microtubules. (A) Shown is localization of mCherry-Radil with eGFP-KIF14 (a–c), mCherry-Radil with eGFP-KIF14-IQAA (d–f), and mCherry-RadilΔPDZ with eGFP-KIF14 (g–i). HEK293T cells were cotransfected with low amounts of Radil expression plasmids together with KIF14 cDNA constructs. (B) Localization of endogenous Radil on microtubules is KIF14 dependent. MDA-MB-231 cells expressing scrambled- Radil or KIF14 shRNA were lysed in BRB80 buffer and cytosolic tubulin was polymerized by addition of 2 mM GTP and 20 µM paclitaxel. Rigor binding of motor proteins was allowed by addition of 2 mM AMP-PNP and the microtubule–motor protein mixture pelleted by centrifugation. The fractionated proteins were detected by Western blotting. P, pellet fraction; S, supernatant fraction.$

KIF14 recruits Radil on microtubules. (A) Shown is localization of mCherry-Radil with eGFP-KIF14 (a–c), mCherry-Radil with eGFP-KIF14-IQAA (d–f), and mCherry-RadilΔPDZ with eGFP-KIF14 (g–i). HEK293T cells were cotransfected with low amounts of Radil expression plasmids together with KIF14 cDNA constructs. (B) Localization of endogenous Radil on microtubules is KIF14 dependent. MDA-MB-231 cells expressing scrambled- Radil or KIF14 shRNA were lysed in BRB80 buffer and cytosolic tubulin was polymerized by addition of 2 mM GTP and 20 µM paclitaxel. Rigor binding of motor proteins was allowed by addition of 2 mM AMP-PNP and the microtubule–motor protein mixture pelleted by centrifugation. The fractionated proteins were detected by Western blotting. P, pellet fraction; S, supernatant fraction.

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