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Figure 2.

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Loss of hamartin (Tsc1) enhances ciliary formation. (A) Western immunoblot of Tsc1−/− and Tsc1+/+ cells showing hamartin levels with β-actin as a loading control. (B and C) MEFs were maintained at confluency for 48 h to induce cilia formation. Cells were 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 the primary cilia. ×100 magnification. (D) Percentage of cells containing a primary cilium is shown, asterisk indicates P < 0.005. (E) Western immunoblot of Tsc1 re-introduction cell lines Tsc1−/−/vector (stably expressing empty vector) and Tsc1−/−/TSC1 (stably expressing TSC1) showing hamartin levels with β-actin as a loading control. Phospho-S6 levels are decreased in cells expressing hamartin, as expected. (F) MEFs were maintained at confluency for 48 h to induce cilia formation. Percentage of cells containing a primary cilium is shown, asterisk indicates P < 0.005. (G) Cells were grown in full serum at subconfluent levels for 24 h. Percentage of cells containing a primary cilium is shown, asterisk indicates P < 0.005.

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