Fig. 3

RAB11 regulates the ciliary import of TTLL5 and TTLL6 in FIP5- and ARL13B-dependent manner. a Depletion of FIP5 or ARL13B impaired ciliary base enrichment for RAB11A-positive vesicles in hTERT-RPE-1 cells. Quantified of RAB11A total intensity at cilia base (30 μm² area were measured). Scale bar: 10 μm. b hTERT-RPE-1 stable cells expressing TTLL5-EYFP and TTLL6-EYFP were treated with 20 nM RAB11A + B siRNAs for 48 h and serum starved for 24 h, the impaired enrichment of TTLL5- or TTLL6-positive vesicles at the ciliary base was observed. Quantified of TTLL5/6-EYFP total intensity at cilia base (30 μm² area were measured). Ac-Tub was immunostained by antibody. Scale bar: 5 μm. c Depletion of ARL13B compromised the enrichment of TTLL5- and TTLL6-positive vesicles at the ciliary base as seen. Quantified of TTLL5/6-EYFP total intensity at cilia base (30 μm² area were measured). Ac-Tub was immunostained by antibodies and TTLL5/6-EYFP was shown by direct fluorescence. Scale bar: 5 μm. d, e Depletion of RAB11 impaired axoneme polyglutamylation (scale bar: 2 μm). Quantification of the length of GT335 signal (d, right panel) and the (GT335 staining)/(Cilia length) ratio (siNC: n = 57, siRAB11A + B: n = 55 cilia) (e) were shown. Center values represent mean. Error bars represent s.d. N value: cilia number accessed from at least six fields. Statistical significance was determined using unpaired Student’s t test. ** p < 0.01