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Fig. 4. Cholesterol-modification is required for Hh homodimerisation. Hh-Np- or Hh-N S2-expressing cells were transfected with an HA-tagged form of Hh-Np (Hh-HA-Np). The presence of the Hh proteins in the culture media or in cell lysates was analysed by western blotting (upper panel). The three different Hh forms are easily distinguishable because of their different electromobility properties. Immunoprecipitation using an anti-HA antibody (lower panel) revealed that Hh-HA-Np co-precipitates with Hh-Np both in cell lysates and in culture media, but does not co-precipitate with Hh-N. The HA adduct imposes a low electromobility onto Hh, whereas the cholesterol adduct in Hh-Np is responsible for its higher electromobility when compared with Hh-N.





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