FIG 4.

GATA3 protein is stabilized in the DN4 subset of Lck-Cre/Fbw7^flox/flox mice. (A) Thymocytes from Fbw7^flox/flox or Lck-Cre/Fbw7^flox/flox mice at 9 to 10 weeks of age were incubated with CD4, CD8, CD25, and CD44 for sorting cells from DN1 to DN4 lineages. A representative gating strategy is shown. The sorted FACS fractions obtained in panel A were subjected to qRT-PCR analysis of Fbw7 expression (B), immunoblot analysis (C), and immunocytochemical staining using Alexa Fluor 546 conjugated with anti-GATA3 antibody (D and E). DAPI staining was also examined for detection of the nuclear location (D and E). In panel C, the arrow and arrowhead indicate Fbw7 and nonspecific signal, respectively.