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. 2009 Sep 21;186(6):817–824. doi: 10.1083/jcb.200906168

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© 2009 Gokhale et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 3. — Biochemical depletion of CK1 rescues microtubule sliding in isolated pf17 axonemes, and this rescue requires I1 dynein. (A) Experimental strategy to test the role of CK1 in microtubules. (B) ATP-induced microtubule sliding velocity was measured in isolated axonemes, CK1-depleted axonemes, or CK1-depleted axonemes reconstituted with purified rCK1 (Okagaki and Kamiya, 1986; Wirschell et al., 2009). The effect of DRB/CK1-7 and the phosphatase inhibitor microcystin-LR (MC) was also examined. The bars represent: (1) wild-type (WT) axonemes; (2) WT axonemes depleted of CK1 (note that there is no change in velocity); (3) pf17 axonemes (note the slow, baseline sliding velocity); (4) pf17 axonemes plus DRB; (5) pf17 axonemes plus CK1-7; (6) pf17 axonemes plus DRB and microcystin-LR; (7) pf17 axonemes plus CK1-7 and microcystin-LR; (8) pf17 axonemes depleted of CK1 (note the rescue of microtubule sliding); (9) pf17 axonemes depleted of CK1 plus microcystin-LR; (10) ida1pf17 axonemes; (11) ida1pf17 axonemes plus DRB; (12) ida1pf17 axonemes plus CK1-7; and (13) ida1pf17 axonemes depleted of CK1 (note the failure in rescue of sliding). Microtubule sliding velocity is expressed as µm/s, and means and standard deviations (error bars) were calculated from at least three independent experiments with a minimum sample size of 75 axonemes.