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. 2011 Jan 27;108(7):2759–2764. doi: 10.1073/pnas.1018823108

Fig. 1.

Fig. 1.

Rab8 localizes to developing cilia but departs mature cilia. (A) Rab8 is an early marker of ciliogenesis. GFP-Rab8a localization in cilia was scored in live RPE GFP-Rab8a cells, and Actub-positive cilia were counted in fixed cells (n = 75 cells). (B) Rab8 accumulates in the growing cilia. One hour after serum withdrawal, RPE GFP-Rab8a cells transiently expressing tRFP-Centrin2 for 24 h were imaged every 10 min by SDC microscopy over 6 h with concurrent imaging in both red and green channels. Representative contrast-adjusted images shown are projections of 20 z-sections (step size 0.5 μm). (Bottom Row) 3D surface rendering. (Scale bar: 5 μm.) (Movie S1). (C) Kinetics of ciliogenesis. Keying on the initiation of nascent cilia, GFP-Rab8a levels in 12 developing cilia were measured from projected z-stacks collected as described in B. Regression analysis was performed on raw data (Inset) to determine the time required for GFP fluorescence to reach maximal levels (106 ± 12 min).