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. Author manuscript; available in PMC: 2014 Feb 7.
Published in final edited form as: Curr Biol. 2012 Sep 13;22(20):1944–1950. doi: 10.1016/j.cub.2012.08.022

Figure 3. Membrane-free Rab11-GTP specifically associates with cenexin at mother centrioles.

Figure 3

(A) Isolated centrosomes from GAPDH- or cenexin-depleted cells were incubated with purified GST-Rab11-GTPγS or GST-Rab11-GDP, spun onto glass coverslips and stained for centrin (green) and GST (red). Below, GST intensity at centrosome was calculated. Representative of n=3 experiments, n>100 centrosomes/condition. P-value is p<1*10-4 between Rab11-GDP and Rab11-GTPγS in control cells, Bar is SE.

(B) Isolated centrosomes were incubated with purified MBP or MBP-Evi5N, spun onto glass coverslips, stained for Rab11 (green), MBP (blue), or centrin. Below, Rab11 intensity at centrosome was calculated. Representative of n=3 experiments, n>30 centrosomes/condition. P-value is p<1*10-4. Bar is SE.

(C) Isolated centrosomes from cells depleted of centriolin were stained for centrin (red), Evi5 (green), and Rab11 (green). Bar, 1 μm. Below, Rab11 (grey) or Evi5 (green) intensity at the centrosome was calculated. Representative of n=3 experiments with p-value <1×10-4, n>50 centrosomes/treatment/experiment, Bar is SE.

(D) Model for regulating Rab11 activity at the mother centriole. Cenexin organizes centriolin, Exocyst, and Evi5. The mother centriole localization of Evi5 can then act on Rab11-GTP and convert it to inactive Rab11-GDP.