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Bodipy-cyclopamine binding assay
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Human Flag Smo WT or Smo mutant (D473H) was transfected in HEK293 cells. 24 h later, the HEK293 cells were trypsinized, washed in phenol-red free DMEM supplemented with 0.5% fetal bovine serum, fixed with 4% paraformaldehyde for 10 min at room temperature and incubated with 5 nM Bodipy-cyclopamine and different concentration of indicated compounds for 2 h at 37 ℃. Then the treated cells were centrifuged and the fluorescent signals were analyzed by flow cytometry.
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