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. Author manuscript; available in PMC: 2013 Apr 20.
Published in final edited form as: Immunity. 2012 Apr 20;36(4):586–599. doi: 10.1016/j.immuni.2012.02.017

Figure 1. I28, F81, and L82 in the N-domains of STAT5A and STAT5B are required for tetramer formation.

Figure 1

(A) Schematic of conserved amino acids required for tetramer binding in STAT1, STAT4, STAT5A, and STAT5B, based on data for STAT1 and STAT4 (Chen et al., 2003). The numbers above (for STAT1 and STAT4) or below (for STAT5A and STAT5B) indicate the amino acid positions. (B through E) EMSAs were performed using [32P]-labeled β-casein (B and C) or IL-2Rα GAScn (D and E) probes and nuclear extracts from 293T+ cells transfected with plasmids directing expression of IL-2Rβ, γc, and JAK3 as well as pCi (B to E, lane 1), pCi-Stat5a (B and D, lanes 2 to 7), or pCi-Stat5b (C and E, lanes 2 to 5), as indicated. Arrows indicate dimer or tetramer binding. (F and G) Off-rates of WT and mutant STAT5 proteins from a site from the bcasein gene, which binds as dimers (F) or a site from IL-2Rα gene, which binds both STAT5 tetramers and dimers (G). (H) Titration of binding WT and mutant STAT5B proteins. The experiments in panel B-H were performed twice.