FIG 5.
ShcFFF expression causes decreased c-Myc expression and impaired Egr1 and Egr3 upregulation. (A) Induction of c-Myc transcript was measured by qPCR after anti-CD3 stimulation of purified DN4 thymocytes from ShcFFF and Lck-Cre/ShcFFF mice (n = 3 experiments, including 9 animals). (B) c-Myc protein expression in DN3, DN4, and DP subsets (n = 3). (C) qPCR analysis of Cdkn1a (p25cip1) and Cdkn1b (p27kip1) mRNA expression in purified DN3E, DN3L, and DN4 thymocytes from Lck-Cre/ShcFFF and control ShcFFF mice (n = 2 with 6 animals per genotype). (D) Induction of Egr1 and Egr3 transcripts was measured by qPCR upon anti-CD3 stimulation of purified DN4 thymocytes from ShcFFF and Lck-Cre/ShcFFF mice (n = 3 experiments with 9 animals per genotype). (E) qPCR analysis of CD3-induced Id3 mRNA expression in DN4 cells from ShcFFF and Lck-Cre/ShcFFF mice (n = 3). (F) Expression of E2A transcript measured by qPCR in CD3-stimulated DN4 thymocytes from ShcFFF and Lck-Cre/ShcFFF mice. (G) Expression of E2A protein measured by intracytoplasmic staining in DN3, DN4, and DP cells from ShcFFF (control) mice and Lck-Cre/ShcFFF mice (n = 2).