Figure 6.
FIP5 acts downstream of CDK6 to regulate axoneme polyglutamylation. (A) The FIP5-mCherry overexpressed RCTE cells were cultured in a serum-containing medium. Images cropped from live-cell imaging before and after Abemaciclib treatment. White triangles indicate the accumulation loci of FIP5-mCherry–positive vesicles. (B) The subcellular localization, and fluorescent intensity of FIP5 at cilia base in indicated YFP-positive RPE-1 cells. Acetylated tubulin (Ac-Tub) was used as a marker of cilium. (C) Knockdown of FIP5 inhibits Abemaciclib-induced axoneme hyperglutamylation in RPE-1 cells. (D) The knockdown efficiency of CDK6 and FIP5 siRNAs was accessed by western blotting. The molecular weigth standards (kD) are labeled on the right. (E) Knockdown of FIP5 inhibits CDK6 depletion-induced axoneme hyperglutamylation in RPE-1 cells. (F and G) Knockdown of FIP5 inhibits CDK6 depletion-induced ciliary import of TTLL5 (F) and TTLL6 (G). Quantified data are presented as mean ± SEM (B, C, E, and right panels of F and G) or SD (left panels of F and G). Statistical analyses were performed by one-way ANOVA analyses with Tukey’s post-hoc test for multiple comparisons. N ≥ 50 cilia (B, C, E, and right panels of F and G) or = 4 independent experiments (left panels of F and G). ***P < 0.001. n.s: not significant. Scale bars: 10 μm (A), 5 μm (B), or 2 μm (C and E–G). Source data are available for this figure: SourceData F6.

FIP5 acts downstream of CDK6 to regulate axoneme polyglutamylation. (A) The FIP5-mCherry overexpressed RCTE cells were cultured in a serum-containing medium. Images cropped from live-cell imaging before and after Abemaciclib treatment. White triangles indicate the accumulation loci of FIP5-mCherry–positive vesicles. (B) The subcellular localization, and fluorescent intensity of FIP5 at cilia base in indicated YFP-positive RPE-1 cells. Acetylated tubulin (Ac-Tub) was used as a marker of cilium. (C) Knockdown of FIP5 inhibits Abemaciclib-induced axoneme hyperglutamylation in RPE-1 cells. (D) The knockdown efficiency of CDK6 and FIP5 siRNAs was accessed by western blotting. The molecular weigth standards (kD) are labeled on the right. (E) Knockdown of FIP5 inhibits CDK6 depletion-induced axoneme hyperglutamylation in RPE-1 cells. (F and G) Knockdown of FIP5 inhibits CDK6 depletion-induced ciliary import of TTLL5 (F) and TTLL6 (G). Quantified data are presented as mean ± SEM (B, C, E, and right panels of F and G) or SD (left panels of F and G). Statistical analyses were performed by one-way ANOVA analyses with Tukey’s post-hoc test for multiple comparisons. N ≥ 50 cilia (B, C, E, and right panels of F and G) or = 4 independent experiments (left panels of F and G). ***P < 0.001. n.s: not significant. Scale bars: 10 μm (A), 5 μm (B), or 2 μm (C and E–G). Source data are available for this figure: SourceData F6.

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