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. Author manuscript; available in PMC: 2014 Jun 16.
Published in final edited form as: Dev Biol. 2011 Sep 16;360(1):66–76. doi: 10.1016/j.ydbio.2011.09.001

Figure 3. Traf3ip1 cells fail to form cilia both in vitro and in vivo and have elevated levels of acetylated α-tubulin in the cytosol.

Figure 3

(A and B) Traf3ip1 GT serum starved mouse embryonic fibroblast (MEF) cells in culture fail to form primary cilia as observed by acetylated α-tubulin immunolabeling compared to Traf3ip1GT MEFs. (B) Note the marked increase in cell size and acetylated α-tubulin levels in Traf3ip1GT mutants, compared to control cells. (C) Total α-tubulin, acetylated α-tubulin, and GAPDH protein levels were determined by western blot and quantitated by densitometry analysis. (D) Traf3ip1 WT and Traf3ip1GT mutant MEF cells were analyzed for cell size/forward scatter (FSC) by flow cytometry. (E and F) For in vivo analysis, E10.5 embryo sections were immunolabeled for the cilia markers Arl13b (red) and acetylated α-tubulin (green) in the lateral plate mesenchyme of a Traf3ip1 WT embryo while there is a near complete loss in this tissue of the Traf3ip1GT mutants. (G and H) Embryonic day 10.5 neural tubes were immunolabeled for acetylated α-tubulin in green with Traf3ip1WT and a Traf3ip1GT mutant. Scale bars 14 μm in A, B and 21μm in C, D and 43μm in G, H. Hoescht nuclear label in blue.