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. 2019 Feb 21;104(8):1597–1607. doi: 10.3324/haematol.2018.192773

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Figure 4. — Identification of the specific domains of RUNX1 and RUNX1-ETO mediating interaction with KLF4. (A) Schematic representation of full-length and truncation mutants of RUNX1 (top) and RUNX1-ETO (bottom). (B and C) FLAG-KLF4 and full-length or truncation mutants of RUNX1 (B) or RUNX1-ETO (C) with MYC tag were co-transfected into 293T cells. At 48 hours (h) after transfection, cell lysates were prepared and underwent immunoprecipitation (IP) with anti-FLAG antibody. Immunoblotting (IB) analyses were performed with anti-MYC antibody. Different combinations of co-transfection with KLF4 and full-length or truncation mutants of RUNX1/RUNX1-ETO were labeled as KLF4/RUNX1, KLF4/RUNX1-ΔRHD, KLF4/RHD, KLF4/RE, KLF4/RE-ΔRHD and KLF4/RHD, respectively. (D) KLF4 and ETO expression plasmids were co-trans-fected into 293T cells. At 48 h after transfection, cells were harvested and cell lysates underwent IP with anti-KLF4 or anti-ETO antibody. IB analysis was performed with the other antibody. WB: western blotting.