ZFIN Image: Liu et al., 2022, Fig. 6.

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Figure Caption

Fig. 6.

AKT1 directly interacts with zEtv2 and zScl. (A,B) AKT1 interacts with zEtv2 and zScl. HEK293T cells transfected with the indicated plasmids were harvested for immunoprecipitation with anti-Flag antibody and western blot analyses. (C) Overexpressed AKT1 interacts with endogenous zEtv2 and zScl. Wild-type embryos were injected with 200 pg Flag-AKT1 mRNA at the one-cell stage, and then harvested for immunoprecipitation with anti-Flag antibody. (D,E) Direct binding of AKT1 to zEtv2 or zScl in vitro. GST, GST-AKT1, GST-zEtv2 (236-345) and GST-zScl were expressed in bacterial cells and purified. The purified GST-fusion proteins GST-zEtv2 (236-345) and GST-zScl were then treated with thrombin to cleave their GST tags, after which they were incubated with GST-AKT1. The pull-down fraction was separated with SDS-PAGE and analyzed by Coomassie Blue staining.

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