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. 2008 Oct 9;18(1):151–163. doi: 10.1093/hmg/ddn325

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Regulation of ciliary formation by Tsc1 and Tsc2 is rapamycin-insensitive. (A) Tsc1^−/−/vector and Tsc1^−/−/TSC1 MEFs were grown in the presence of 10% FBS for 48 h with rapamycin (20 nmol) or DMSO control. Percentage of cells containing a primary cilium is shown, asterisk indicates P < 0.005, P = NS, no significant difference. (B) Tsc2^−/−p53^−/−and Tsc2^+/+p53^−/− MEFs were grown in the presence of 10% FBS for 48 h with rapamycin (20 nmol) or DMSO control. Percentage of cells containing a primary cilium is shown, asterisk indicates P < 0.005, P = NS, no significant difference. (C and D) Tsc2^−/−p53^−/− and Tsc2^+/+p53^−/− MEFs were serum starved for 72 h in the presence of rapamycin (20 nmol) or DMSO control. (C) Percentage of cells containing a primary cilium is shown. (D) Percentage of cells containing more than one primary cilium is shown, asterisk indicates P < 0.005, section sign indicates P < 0.05, P = NS, no significant difference. (E–G) Examples are shown of single Tsc2^−/−p53^−/− cells containing more than one primary cilium. MEFs were serum starved for 72 h, fixed and immunostained with anti-α-acetylated tubulin antibody (cilia, green) and anti-γ-tubulin antibody (centrosomes/basal body, red). Nuclei were stained with DAPI. Arrows indicate primary cilia. ×100 magnification.