FIGURE 4. Collagen binding to TRα2[I332G]β1-Fc cation-binding site mutants.
Culture supernatants from transfected CHO L761h cells with TRα2 and wild-type TRβ1 or TRβ1 mutated at the MIDAS (D130A), ADMIDAS (D137A, D138A) or LIMBS (D226A, E169A, E169D) were analysed in Fc-capture assays, performed in the presence of 5mM Mg2+ (A) or 1 mM Mn2+ (B). The ligand-binding capacity of the α2β1-Fc mutants was detected with 0.5 μg/ml collagen I in the absence (open bars) or the presence of anti-β1 antibody TS2/16 (diagonally hatched bars). Absorbance values and error bars represent the mean and standard deviation from one experiment, representative of three separate experiments.