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. 2016 Mar 1;27(5):838–847. doi: 10.1091/mbc.E15-10-0707

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© 2016 Meng and Pan. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — cnk4 flagella exhibit normal IFT speed but reduced IFT frequency, with frequent IFT stalling. (A) Expression of yellow fluorescent protein (YFP)–tagged IFT46 in cnk4. Cells were analyzed by immunoblotting with antibodies against IFT46, GFP, and α-tubulin. (B) Kymographs of IFT46-YFP in the flagella of ift46::IFT46-YFP (a) and cnk4::IFT46-YFP (b–d) cells. Left, images of recorded flagella. (b) Kymograph from apparently normal flagellum. The arrows and arrowhead indicate stalled IFT in anterograde and retrograde directions, respectively. (c) Kymograph highlighting IFT turnaround at the bulged flagellar tip. Arrow, spontaneously initiated retrograde IFT. (d) Kymograph showing IFT at the flagellar bulge in the middle of the flagellum. Arrow, trapped IFT; arrowhead, IFT passing though the bulge. B, flagellar base; T, flagellar tip. Bars, 1 μm and 1 s. (C) The velocities of anterograde and retrograde IFT are not affected by cnk4 mutation. n.s., not significant (p > 0.05). (D) IFT frequencies of both directions in cnk4 flagella are reduced compared with those of wild-type flagella.