JNK is closely associated with the BBs of both motile and primary cilia. (A) Immunostaining of stage 32 Xenopus embryos with anti-phospho-JNK (pJNK) and anti-acetylated-α-tubulin (ac Tubulin) to visualize cilia. pJNK is localized at the base of motile cilia. Scale bars, 10 μm. (B–B’’) Higher magnification images of pJNK at the base of cilia. Scale bars, 1 μm. (C) GFP-JNK and RFP-Centrin mRNA were injected into the ventral marginal region of two blastomeres at the four-cell stage, and the embryos were observed at stage 32. JNK forms a ring around basal bodies Scale bars, 5 μm. (D) GRP ciliated cells of a stage 17 embryo expressing RFP-Centrin and GFP-JNK1, immunostained for acetylated tubulin. GFP-JNK1 is associated with the basal bodies in GRP ciliated cells. Scale bar, 5 μm. (E) Neural tube cross section of a stage 22 embryo coexpressing GFP-JNK1 and centrin2 RFP and immunostained for acetylated tubulin. Scale bar, 5 μm. (E’) The right panels represent magnified insets marked by white squares. Scale bar, 2 μm. (F) Confocal immunofluorescence images of serum-starved HeLa cells showing that pJNK is localized at the base of primary cilia. Scale bar, 2 μm. (G) Immunofluorescent images of human nasal MCCs showing anti-acetylated tubulin and anti-pJNK showing that pJNK is localized at the base of motile cilia. Scale bar, 2 μm. (H) Immunofluorescent images of human nasal MCCs showing anti-acetylated tubulin and anti-tJNK. Scale bar, 2 μm. (I–I’’) Representative confocal images of endogenous localization of acetylated tubulin and JNK (tJNK) in mouse tracheal MCCs showing apical enrichment of JNK. Scale bar, 5 μm. (J) Western blot analysis showing elevated pJNK and JNK protein levels in skin lysates from embryos overexpressing multicilin and controls. Quantification shows the normalized ratio of pJNK/GAPDH intensity. Data represent mean ± SEM from three independent experiments. ***, P < 0.001, unpaired two-tailed t test. (K) Optical section of an intercalating multiciliated cell expressing GFP-JNK1 and RFP-Centrin and stained for actin. GFP-JNK1 is associated with the basal bodies during the early stages of ciliogenesis. Scale bar, 20 μm. Source data are available for this figure: SourceData F1.
Figure 1.

JNK is closely associated with the BBs of both motile and primary cilia. (A) Immunostaining of stage 32 Xenopus embryos with anti-phospho-JNK (pJNK) and anti-acetylated-α-tubulin (ac Tubulin) to visualize cilia. pJNK is localized at the base of motile cilia. Scale bars, 10 μm. (B–B’’) Higher magnification images of pJNK at the base of cilia. Scale bars, 1 μm. (C) GFP-JNK and RFP-Centrin mRNA were injected into the ventral marginal region of two blastomeres at the four-cell stage, and the embryos were observed at stage 32. JNK forms a ring around basal bodies Scale bars, 5 μm. (D) GRP ciliated cells of a stage 17 embryo expressing RFP-Centrin and GFP-JNK1, immunostained for acetylated tubulin. GFP-JNK1 is associated with the basal bodies in GRP ciliated cells. Scale bar, 5 μm. (E) Neural tube cross section of a stage 22 embryo coexpressing GFP-JNK1 and centrin2 RFP and immunostained for acetylated tubulin. Scale bar, 5 μm. (E’) The right panels represent magnified insets marked by white squares. Scale bar, 2 μm. (F) Confocal immunofluorescence images of serum-starved HeLa cells showing that pJNK is localized at the base of primary cilia. Scale bar, 2 μm. (G) Immunofluorescent images of human nasal MCCs showing anti-acetylated tubulin and anti-pJNK showing that pJNK is localized at the base of motile cilia. Scale bar, 2 μm. (H) Immunofluorescent images of human nasal MCCs showing anti-acetylated tubulin and anti-tJNK. Scale bar, 2 μm. (I–I’’) Representative confocal images of endogenous localization of acetylated tubulin and JNK (tJNK) in mouse tracheal MCCs showing apical enrichment of JNK. Scale bar, 5 μm. (J) Western blot analysis showing elevated pJNK and JNK protein levels in skin lysates from embryos overexpressing multicilin and controls. Quantification shows the normalized ratio of pJNK/GAPDH intensity. Data represent mean ± SEM from three independent experiments. ***, P < 0.001, unpaired two-tailed t test. (K) Optical section of an intercalating multiciliated cell expressing GFP-JNK1 and RFP-Centrin and stained for actin. GFP-JNK1 is associated with the basal bodies during the early stages of ciliogenesis. Scale bar, 20 μm. Source data are available for this figure: SourceData F1.

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