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>