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. 2021 Feb 22;10:600762. doi: 10.3389/fonc.2020.600762

Figure 4.

Figure 4

MLK3 directly binds with EPS8. The results shown are representative of at least three independent experiments. (A, B) Overexpressed and endogenous MLK3 binds to EPS8 in U251 cells. U251 cells were cotransfected with pcDNA3.1-Myc-MLK3 and pcDNA3.1-His-EPS8 plasmids for 24 h, with the pcDNA3.1 plasmid as a control. The lysates were immunoprecipitated using anti-Myc (A), anti-EPS8 (B) or control IgG, EPS8, or MLK3 was detected with an immunoblot assay. (C) The molecular structure of MLK3 and EPS8. (D) AAs 1–104 and 632–847 of MLK3 directly bound to EPS8 in vitro. GST-tagged MLK3 segments were expressed in BL21 cells. The purified proteins were incubated with the protein lysates from pcDNA3.1-EPS8-transfected HEK293 cells. The proteins were immunoblotted with indicated antibodies. * the target protein. (E) The expression of MLK3 and EPS8 predicts the overall survival of patients. The data were derived from the CGGA dataset (n = 286). Kaplan-Meier survival analyses for MLK3 and EPS8 mRNA levels. Log-rank test. *P < 0.05, **P < 0.01.