Fig. 2.

Skp2 interacts with FOXO1 in vivo and in vitro. LNCaP cells were transfected with the indicated plasmids, and protein lysates were immunoprecipitated (IP) with anti-FLAG antibody (M2) (A) or an anti-HA antibody (12CA5) (B). A mouse IgG was used as a negative control. Immunoprecipitates were analyzed by immunoblotted (IB) with anti-FLAG and anti-HA antibodies. (C) Protein lysates from NIH 3T3 cells were immunoprecipitated with an anti-Skp2 antibody and immunoblotted with antibodies for FOXO1 and Skp2. (D) Lysates of NIH 3T3 cells transfected with FLAG-FOXO1 were subjected to GST pull-down by GST-Skp2 purified from bacteria.