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. 2019 Apr 12;8(4):28. doi: 10.1038/s41389-019-0138-y

Fig. 4. ZFHX3 and ERβ physically interact with each other independent of DPN treatment.

Fig. 4

a Cell lysates from C4-2B cells, which express both ZFHX3 and ERβ, were subjected to immunoprecipitation (IP) with anti-ERβ antibody and subsequent immunoblotting (IB) with anti-ZFHX3 antibody. Input indicates cell lysate not subjected to IP. b C4-2B cells were grown in phenol red-free medium with 5% charcoal-stripped FBS for 48 h and then treated with DPN for 48 h. IP and IB were performed as in a. c Expression plasmids for HA-tagged ZFHX3 (HA-ZFHX3) and FLAG-tagged ERβ (FLAG-ERβ) were transfected into 293T cells. Lysates were subjected to IP with anti-FLAG or anti-HA affinity gel, and then to IB with anti-HA or anti-FLAG antibody. d Schematic of full ZFHX3 protein (3703 residues, horizontal bar) with 23 zinc fingers (gray ovals) and 4 homeodomains (black rectangles). The six shorter bars below indicate six overlapping fragments of ZFHX3, named A to F. Each of the six fragments was tagged with HA, expressed in 293T cells, and tested for their interactions with ERβ by IP and IB. The two confirmed interactions with ERβ, A and D, are shown in solid dark. e IP and IB results for the interaction of FLAG-tagged ERβ and each of the six HA-tagged ZFHX3 fragments. The same procedures as in c were used. Arrows indicate the two fragments that were pulled down by ERβ (i.e., a and d). ZFHX3, zinc-finger homeobox 3; ERβ, estrogen receptor beta; FBS, fetal bovine serum; DPN, diarylpropionitrile