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. 2017 Aug 29;67(1):46–57. doi: 10.2337/db17-0546

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© 2017 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. More information is available at http://www.diabetesjournals.org/content/license.

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SCP4 physically interacts with FoxO1/3a. A: SCP4 co-IP with FoxO1. HEK293T cells were transfected with FLAG-FoxO1 and HA-SCP4. Co-IP was carried out by using an anti-FLAG antibody. Levels of indicated proteins in whole cell lysates (WCL) and IP products were analyzed by IB. B: SCP4 co-IP with FoxO3a. HEK293T cell transfection, co-IP, and IB were done as described in A. C: SCP4 interacts with FoxO1/3a at endogenous levels in vivo. Cell lysates of HepG2 cells were IP with an anti-SCP4 antibody or control IgG antibody. The immunocomplexes and input were analyzed by IB with indicated antibodies. D: SCP4 directly binds to FoxO1 in vitro. In vitro binding assay was carried out with purified GST-SCP4 absorbed on Glutathione Sepharose beads and in vitro translated FoxO1. IB was carried out with indicated antibodies. E: SCP4 directly binds to FoxO3a in vitro. The in vitro binding assay and IB were carried out as described in D.