Impaired interaction of mutant protein with the IKAROS family. (A-C) 293T cells were transfected with Flag-tagged WT IKAROS, WT AIOLOS, WT HELIOS, or hemagglutinin (HA)-tagged WT IKAROS or mutants (R213*, S427*, C467R, R502L, Y462* [a laboratory-generated mutant with deletion of ZF5 and ZF6]). Immunoprecipitation (IP) was performed using anti-Flag antibodies. Western blot analysis of the IP samples with anti-HA and anti-Flag or AIOLOS or HELIOS antibodies is shown. Five percent of the total volumes of the whole-cellular lysates used for each IP reaction were loaded as input controls. Data are representative of 3 independent experiments. Densitometry analysis of immunoprecipitated HA-IKAROS blots was performed using ImageStudio. EV, empty vector control.