Figure 1.
FKBP51 is modified by SUMO conjugation. (a) FKBP51 was SUMOylated in vitro and analyzed by western blotting using anti-FKBP51 antibody. (b) HEK293T cells were transfected with the indicated plasmids. SUMOylated proteins were purified by Ni2+ affinity chromatography and analyzed by western blotting using anti-FKBP51 antibody. (c) HEK293T cells were transfected with the indicated plasmids and analyzed as in (b). (d) Pull-down assays were performed using HEK293T lysates expressing FLAG-FKBP51 and analyzed by western blotting using anti-FKBP51 antibody. (e) HEK293T lysates expressing FLAG-FKBP51, HA-SUMO2 and V5-Ubc9 were immunoprecipitated with anti-FKBP51 antibody, and immunoprecipitates were incubated with or without recombinant SENP2. Samples were analyzed by western blotting using anti-FKBP51 antibody. (f) HEK293T cells were transfected with the indicated plasmids and analyzed as in (b). (g) Lysates from HEK293T were immunoprecipitated with anti-SUMO2/3 antibody (ab) or non-immune antibody (NI ab) and analyzed by western blotting using the indicated antibodies. *Non-specific. (h) HT22 cells were transfected with the indicated plasmids and analyzed as in (b)