10.1371/journal.pone.0113839.g005 Chung Hyun Park Chung Hyun Park Danalea V. Skarra Danalea V. Skarra Alissa J. Rivera Alissa J. Rivera David J. Arriola David J. Arriola Varykina G. Thackray Varykina G. Thackray FOXO1 Interacts with SMAD3/4. Public Library of Science 2014 activin induction FOXO 1 FOXO 1 Diminishes Activin Induction FOXO 1 suppression FOXL L βT cells FOXO 1 decreases activin induction FOXO 1 binding smad activin response elements FOXO 1 transcription factor modulates activin FOXO 1 DNA binding FOXO 1 interaction Fshb promoter activin induction maps dbd murine Fshb gene expression 2014-11-25 02:43:57 Figure https://plos.figshare.com/articles/figure/_FOXO1_Interacts_with_SMAD3_4_/1251341 <p>A. GST interaction assays were performed using bacterially expressed GST-fusion proteins (indicated above each lane) and <sup>35</sup>S-labeled <i>in vitro</i> transcribed and translated GFP, FOXO1, FOXO1-CA, and FOXO1-CA-DBD mutant (indicated on the left of the panel). GFP was used as a negative control. The GST-fusion proteins included GST alone, GST-SMAD2 (S2), GST-SMAD3 (S3), and GST-SMAD4 (S4). One quarter of the protein used in the interaction assay was loaded in the lane marked input. The experiment was repeated several times with similar results and a representative experiment is shown. B. Co-immunoprecipitation assays were performed using nuclear extracts from LβT2 cells treated with or without 10 ng/mL activin for 2 h after an overnight incubation in serum-free media. One tenth of the protein used in the immunoprecipitation reaction was loaded in the lane marked input. The immunoprecipitation was performed with mouse IgG, SMAD4 (S4) or SMAD2/3 (S2/3) antibody, as indicated. Western blot analysis was performed using SMAD4, SMAD2/3 and FOXO1 primary antibodies, and a horseradish peroxidase-linked secondary antibody. The experiment was repeated several times with similar results and a representative experiment is shown.</p>