Skip to main content
. Author manuscript; available in PMC: 2013 Aug 7.
Published in final edited form as: Oncogene. 2012 Mar 5;32(3):388–396. doi: 10.1038/onc.2012.63

Figure 2. Mapping the MDM2-RPS14 binding domains.

Figure 2

(A) RPS14 directly binds to the central acidic domain of MDM2 in vitro. Purified GST, GST-MDM2, or GST-MDM2-fragments immobilized on glutathione beads were incubated with 500 ng of bacterially purified His-RPS14. Bound proteins were blotted with anti-His. Arrows indicate the GST-MDM2 or GST-MDM2-fragments. (B) RPS14 binds to the central acidic domain of MDM2 in cells. Cells were transfected with Flag-RPS14, V5-MDM2, or deletion mutants of V5-MDM2 plasmids and harvested for IP (0.5 mg total proteins) with anti-V5, followed by IB with the antibodies as indicated. Arrows indicate the V5-MDM2, V5-MDM2-fragments or heavy chains (HC). (C) MDM2 binds to the C-terminus of RPS14 in cells. Cells were transfected with HA-MDM2, Flag-RPS14, or deletion mutants of Flag-RPS14 plasmids and harvested for IP (0.5 mg total proteins) with anti-Flag, followed by IB with the antibodies as indicated. (D) Schematic presentation of the MDM2 domain(s) that binds to RPS14. (E) Schematic presentation of the RPS14 domain(s) that binds to MDM2.