Fig. 6
Free NHR2 domain of CBFA2T3 inhibits the cooperative effect of CBFA2T3 on RUNX1 activity. a–d Luciferase assays with the RUNX1-consensus motif repetition (a), the RUNX1 promoter (b), the CBFA2T3 enhancer (c) and the c-KIT enhancer (d) upstream a minimal promoter and a luciferase ORF, in presence of RUNX1 and CBFA2T3 or CBFA2T3NHR2 expressing plasmids in HEK293 cells. Luciferase levels (Firefly luciferase/Renilla luciferase) are represented using a scatter dot plot indicating the means and S.D. ns: non-significant, **p < 0.01, ***p < 0.001, ****p < 0.0001 in Mann–Whitney tests compared to the control condition. e Relative mRNA expression of RUNX1, CBFA2T3 and c-KIT measured by RT-qPCR in REH cells, and REH expressing CBFA2T3NHR2 cells (REH+NHR2). Results are presented in-terms of a fold change after normalizing with ABL mRNA. f Schematic representation of the activation loop between RUNX1 and CBFA2T3. Expression of the free NHR2 domain interrupts this loop