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Figure 7


Fig. 7. Ciliogenesis and Shh signalling in Ftm-/- cells in mice. (A) Upper part: immunohistochemistry on G0 phase (left) and proliferating (right) MLCs. (Left panels) In G0 phase, each wild-type and Ftm-/- cell exhibits a single cilium, detected by acetylated {alpha}-tubulin antibody (green staining). Ftm (stained in red) is present only in wild-type MLCs. (Right panels) In proliferating wild-type and Ftm-/- MLCs, cilia are absent and the cytoplasm is enriched with cytoskeletal tubulins, marked by acetylated {alpha}-tubulin antibody (green staining). Note that Ftm antibodies were not used in this staining. Lower part: Shh response in G0-phase (left) and proliferating (right) wild-type and Ftm-/- MLCs. G0-phase wild-type MLCs show a strong induction of Gli1 and Ptc1 after incubation with recombinant Shh protein for 24 hours. In G0-phase Ftm-/- MLCs, the induction of Gli1 and Ptc1 is strongly reduced. Both, proliferating (non-ciliated) wild-type and Ftm-/- MLCs show no induction of Shh target genes after stimulation. (B) Quantification of Ptc1-expression levels in G0-phase (left panels) and proliferating (right panels) wild-type and Ftm-/- MLCs after Shh stimulation. The quantification of Ptc1 induction reveals that G0-phase wild-type MLCs show an increase of Ptc1 transcripts by a factor of 9.4, whereas G0-phase Ftm-/- MLCs show an increase of only a factor of 3.1. Proliferating (non-ciliated) wild-type and Ftm-/- MLCs show no changes in the relative expression levels of Ptc1 upon treatment with Shh (wild-type MLCs 1.1; Ftm-/- MLCs 0.8). Data represent the quantification of three independent experiments. wt, wild type.





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