Fig. 4.

LR inhibits GMR complex formation. (A) The 293T cells were transfected with αGMR plus βGMR (2.5 μg each) or 2.5 μg of αGMR plus 2.5 μg of βGMR plus 5 μg of LR. After transfection (48 h), cells were harvested and lysed; the lysates were then cleared by centrifugation. The lysates were treated with 0.5 nM GM-CSF for 12 h at 4°C. Immunoprecipitation (IP) was done with βGMR Ab S16. In Western blot, αGMR was detected by C18 Ab and the immunoprecipitated βGMR was detected by Ab C20. (B) Total protein levels of αGMR and LR in lysates used for the immunoprecipitation experiment (A) were detected by C18 Ab and anti-V5-HRP, respectively.