Fig. 2.
Ligand binding induces mono-/polyubiquitination of the G-CSFR. ts20 cells expressing V5-tagged WT G-CSFR were incubated at 37°C with G-CSF (100 ng/mL) for the indicated times, lysed, then immunoprecipitated with anti-V5 antibody and blotted with the indicated antibodies. The FK1 antibody (A) recognizes polyubiquitinated proteins whereas the FK2 antibody (B) recognizes both poly- and mono-ubiquitinated proteins. (C) The blot was stripped and re-blotted with anti-V5 antibody. Histograms are shown to the right of each blot depicting the density ratio between bands of FK1 and FK2 to anti-V5, respectively, as a control for receptor protein loading.
