Skip to main content
. 2019 Apr 16;10:568. doi: 10.3389/fimmu.2019.00568

Figure 2.

Figure 2

(A) The sequence, used for the EMSA experiments is shown. Motifs that resemble the IKAROS consensus are shown in red in the rectangle. (B) Mutations affecting the DNA binding domain of IKAROS, preventing its binding to its target sequence. A specific gel-shift band, indicating binding of IKAROS multimers to the IKBS-4 DNA probe, was observed in the nuclear extract of HEK293T cells expressing the wildtype IKAROS. This band was absent in cells expressing the IKAROS variants Arg143Trp, Cys150Arg, and Arg162Gln and was weaker in the nuclear extracts of cells co-transfected with equal amounts of wildtype and one of the three mutant forms. A smaller band was also detected in the co-transfected samples, compared to the wildtype ones which can indicate the DNA-bound dimers of IKAROS. The DNA binding activity remained unaffected when the mutant variants Lys286*, Gly337Ser, Ala448Arg, and Met494Val were expressed.