Figure 8: LIMK2 and TESK1 control vesicle trafficking to the pericentrosomal area.

(a) Quantification of ciliogenesis in RPE1 cells transfected with the indicated siRNAs for 60 h. Cells were serum-starved for final 24 h. (b) Graph showing the measurement of ciliary length in cells treated with serum starvation (24 h), CytoD (24 h) or YAP/TAZ mix siRNAs (60 h). (c) Immunofluorescence images showing the effect of LIMK2 and TESK1 knockdown on the trafficking of tentative ciliary vesicles double-positive for Smo-EGFP and endogenous RAB11. CEP164 was also depleted to arrest ciliogenesis at the initiation stage of ciliogenesis. Insets are magnified view of the centrosomal area. (d) Quantification of experiments illustrated in c. (e) Subcellular localization of endogenous LIMK2 and flag-tagged TESK1, which was transiently expressed. Insets are magnified view of the centrosomal area. (f) A proposed model for the regulation of cilium assembly by actin-dependent pathways. All scale bars are 10 μm. All error bars are s.d. (n=3 independent experiments). The P-values were determined by unpaired Student’s t-test; **P<0.01.