Expression of GATA2-mutants in HEK 293T cells. (A) Expression of FLAG-tagged GATA2-mutants confirmed by 3 independent Western blot (WB) experiments using anti-GATA2 antibodies (top) and anti–β-actin antibodies (bottom). Graph shows mean values of the GATA2 signals normalized to β-actin signals as fold change compared with wild-type GATA2. Error bars indicate standard error of the mean. (B) Immunofluorescence of GATA2 using anti-FLAG antibodies and Alexa 555–labeled secondary antibodies. DNA was counter stained using 4,6-diamidino-2-phenylindole. Confocal laser scans show nuclear localization for GATA2 wild type (top) and the recurring GATA2-mutants A318T (middle) or G320D (bottom). Similar results were obtained for all GATA2-mutants tested (see supplemental Figure 3).