Words similar to kaad-cyclopamine
Example sentences for: kaad-cyclopamine
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Binding was assayed in the presence of KAAD-cyclopamine at 10 μM (Figure 6c, column 4), tomatadine at 10 μM (Figure 6c; Antag control 1, column 5), Cur61414 at 10 μM (Figure 6c, column 6), or the inactive Cur61414 derivative (Figure 6c; Antag control 2, column 7) at 10 μM.
We next compared binding of KAAD-cyclopamine, Cur61414 and Hh-Ag 1.5 to a constitutively active mutant of Smo (Smo act) or to wild-type Smo (Smo wt).
Using Smo act- and Smo wt-containing membranes, we then performed competition binding studies by adding increasing concentrations of unlabeled Hh-Ag 1.5, KAAD-cyclopamine or Cur61414 in the presence of [ 3H]-Hh-Ag 1.5 (0.
To perform theses studies, Smo-overexpressing 293T cells were incubated for 2 hours at 37°C with 5 nM [ 3H]-Hh-Ag 1.5 in the presence of either KAAD-cyclopamine at 5 μM (Figure 6a, column 6), the related but inactive plant compound tomatadine at 5 μM (Figure 6a; Antag control 1, column 7), Cur61414 at 5 μM (Figure 6a, column 8), or a related but inactive Cur61414 derivative (Figure 6a; Antag control 2, column 9) at 5 μM.
The two Hh-signaling antagonists, KAAD-cyclopamine and Cur61414, have shown decreased potency on Smo act-expressing cells, leading to the speculation that they may bind this mutant form of Smo less well than the wild-type form [ 11 17 ] . We sought to determine whether the Hh agonist binds Smo actwith a higher affinity, an observation seen with certain ligands and constitutively active mutants of GPCRs [ 29 ] . To perform this experiment, we isolated membranes from cells transfected with a cDNA construct encoding a tryptophan-to-leucine mutation at residue 539 (W539L) of murine Smo.
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