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. 2016 Feb 1;30(3):281–292. doi: 10.1101/gad.274118.115

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© 2016 Coffill et al.; Published by Cold Spring Harbor Laboratory Press

This article, published in Genes & Development, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 7. — Lj-Mdm2 function and binding partners are conserved across species. (A) In vitro pull-down assay of Hs-p53 or Lj-p53 by HDM2^Nterm or Lj-Mdm2^Nterm in the presence of Nutlin or Staplin. (D) DMSO; (N) Nutlin; (S) Staplin. The top panel shows immunoprecipitated p53, and the bottom panel shows input levels. Hs-p53 was detected using DO1 antibody, and Lj-p53 was detected using anti-Flag antibody. (B) Western blot of Hs-p53 levels following cotransfection with an Lj-Mdm2-expressing construct. (Lane 1) Flag-Hs-p53. (Lane 2) Flag-Hs-p53 with MG132. (Lane 3) Flag-Hs-p53 + HA-Lj-Mdm2. (Lane 4) Flag-Hs-p53 + HA-Lj-Mdm2 with MG132. The loading control is shown in the bottom panel. (C, top panel) Western blot of in vitro translation and immunoprecipitation (IP) of Hs-RPL5 or Hs-RPL11 by either HDM2 or Lj-Mdm2, which was used as bait. (Lane 1) Hs-RPL5 with HDM2. (Lane 2) Hs-RPL5 with Lj-Mdm2. (Lane 3) Hs-RPL5 with blank. (Lane 4) Hs-RPL11 with HDM2. (Lane 5) Hs-RPL11 with Lj-Mdm2. (Lane 6) Hs-RPL11 with blank. Input levels are shown in the bottom panel.