Fig. 3.
Oncogenic fusion to FAM22 enables aberrant nuclear localization of YWHAE. (A) Endogenous YWHAE–FAM22A is predominantly nuclear, whereas native YWHAE is predominantly cytoplasmic. FOXO3A and poly(ADP-ribose) polymerase (PARP) are nuclear localization controls, whereas GAPDH is a cytoplasmic control. (B) Induced YWHAE–FAM22A expression is nuclear in 293T cells, as shown by FLAG immunoprecipitation (Upper) and YWHAE immunohistochemistry (Lower) after transient expression of FLAG-tagged YWHAE–FAM22A pcDNA3 construct. In contrast to the predominantly cytoplasmic staining (and absent nuclear staining) seen in nontransfected 293T cells (representing wild-type YWHAE), YWHAE immunostaining in YWHAE–FAM22A–expressing 293T cells showed the presence of nuclear staining, indicating nuclear localization of the fusion protein.