Primary Cilia Are Not Required for Normal Canonical Wnt Signaling in the Mouse Embryo (original) (raw)
Figure 4
The response of fibroblasts to Wnt3a does not depend on cilia.
A. Wild type, Ift88, Ift172 and Dync2h1 MEFs stained for cilia (acetylated α-tubulin, green) and basal bodies (centrin, red). MEFs derived from e9.5 wild type and Dync2h1 embryos generate cilia within 24 hours of culture, but not Ift172 or Ift88 mutant MEFs. B. Ift88, Ift172 and Dync2h1 MEFs fail to respond to Shh. Cells were transfected with a Hh-responsive Gli-luciferase reporter and stimulated with Shh-enriched media. Wild-type cells showed robust activation of the reporter in response to Shh treatment, whereas Ift88, Ift172 and Dync2h1 MEFs were completely non-responsive to Shh. C. Ift88, Ift172 and Dync2h1 MEFs respond normally to Wnt3a. SuperTOP-Flash reporter activity was assayed in response to 100 ng/mL recombinant Wnt3a. Reporter activity presented as relative light units (RLU) normalized to Renilla luciferase control. Ift172 and Dync2h1 mutant MEFs activated the reporter in response to different levels of Wnt3a similar to wild type levels (data are mean ±s.d., n = 4).