JNK is involved in the association of IFT-52 with the basal bodies and apical actin network establishment. (A) (I) Representative images of embryos injected with GFP-CEP83 and RFP-Centrin in control and JNK MO embryos. (II) Images of embryos injected with GFP-CEP123 and RFP-Centrin in control and JNK MO embryos. (III) Control and JNK morphants stained with Phalloidin and CEP164. (IV) Representative image of MCC injected with mCherry-IFT46 and stained for Centrin. The association of CEP83, CEP123, CEP164 and IFT46 with the basal bodies is not affected in JNK morphants. Scale bars, 10 µm. (B) Representative confocal images of stage 19 embryos coinjected with GFP-IFT52 and RFP-Centrin. IFT52 is associated with the basal bodies during the early stages of ciliogenesis. Scale bars, 5 µm. (C) MCCs of DMSO- and SP600125-treated embryos stained with phalloidin to visualize apical and subapical actin networks. Single optical sections show impaired apical and subapical actin networks in SP600125-treated embryos. Scale bars, 5 µm. (D) Quantification of the half-time of recovery (t1/2) of apical actin in control and JNK morphant MCCs. Error bars indicate ± SEM. ***, P < 0.001, unpaired two-tailed t test. (E) Optical sections of a Xenopus epidermal MCC expressing GFP-JNK and RFP-Utrophin. A second region of JNK enrichment in close association with the subapical actin is occasionally detected. Scale bars, 5 μm. (F) Multiciliated cell expressing GFP-JNK (donor) and mKate2 Actin (acceptor) showing the apical actin network, before and after acceptor photobleaching (n = 5 from 3 embryos). GFP intensity rises, showing that FRET is taking place between GFP and mKate2 suggesting that JNK interacts with actin. Scale bars, 10 μm.
Figure S4.

JNK is involved in the association of IFT-52 with the basal bodies and apical actin network establishment. (A) (I) Representative images of embryos injected with GFP-CEP83 and RFP-Centrin in control and JNK MO embryos. (II) Images of embryos injected with GFP-CEP123 and RFP-Centrin in control and JNK MO embryos. (III) Control and JNK morphants stained with Phalloidin and CEP164. (IV) Representative image of MCC injected with mCherry-IFT46 and stained for Centrin. The association of CEP83, CEP123, CEP164 and IFT46 with the basal bodies is not affected in JNK morphants. Scale bars, 10 µm. (B) Representative confocal images of stage 19 embryos coinjected with GFP-IFT52 and RFP-Centrin. IFT52 is associated with the basal bodies during the early stages of ciliogenesis. Scale bars, 5 µm. (C) MCCs of DMSO- and SP600125-treated embryos stained with phalloidin to visualize apical and subapical actin networks. Single optical sections show impaired apical and subapical actin networks in SP600125-treated embryos. Scale bars, 5 µm. (D) Quantification of the half-time of recovery (t1/2) of apical actin in control and JNK morphant MCCs. Error bars indicate ± SEM. ***, P < 0.001, unpaired two-tailed t test. (E) Optical sections of a Xenopus epidermal MCC expressing GFP-JNK and RFP-Utrophin. A second region of JNK enrichment in close association with the subapical actin is occasionally detected. Scale bars, 5 μm. (F) Multiciliated cell expressing GFP-JNK (donor) and mKate2 Actin (acceptor) showing the apical actin network, before and after acceptor photobleaching (n = 5 from 3 embryos). GFP intensity rises, showing that FRET is taking place between GFP and mKate2 suggesting that JNK interacts with actin. Scale bars, 10 μm.

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