Figure 3.

IKAROS subnuclear and pericentromeric-heterochromatin localization. (A) NIH3T3 cells, transiently transfected with the pCR3 vector expressing wildtype or mutant IKAROS were incubated with rabbit-anti-human-IKAROS antibody and N-terminus anti-human IKAROS antibody and stained with goat anti-rabbit Alexa fluor 488. Cells overexpressing the wildtype IKAROS, showed the punctate staining pattern, observed in green. In contrast, a diffuse nuclear staining was observed in NIH3T3 cells overexpressing the Arg143Trp, Cys150Arg, Arg162Gln, and Lys286* mutant forms. (B) The DNA binding ability of wildtype IKAROS was not inhibited by the presence of a mutant variant. NIH3T3 cells were co-transfected with equal amounts of wild-type and each mutant pCR3 expression vectors and incubated and stained with the above-mentioned antibodies. In NIH3T3 cells co-transfected with equal amounts of wild-type and each mutant forms, the punctate nuclear staining pattern were obtained in all samples.