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. 1997 Dec 9;94(25):13850–13855. doi: 10.1073/pnas.94.25.13850

Figure 5.

Figure 5

Regulation of STAT5A DNA binding by mutation of JAK3 residues Y980 and Y981. COS-7 cells were transfected with 5 μg STAT5A cDNA only (lanes 1, 8, 13, and 17) or cotransfected with 5 μg STAT5A and 5 μg cDNAs encoding: WT JAK3 (lanes 2, 9, 14, and 18), K855A (lane 3), Y980F (lanes 4, 10, 15, and 19), Y981F (lanes 5, 11, 16, and 20), or YY980/981FF (lanes 6 and 12), or mock transfected (lane 7). Two days later, cells were harvested and nuclear extracts were prepared. Protein determination was measured using BCA protein assay reagent (Bio-Rad). EMSA was performed using 5 μg protein and a labeled probe from the GAS-like element in the CD23 promoter (lanes 1–7). Competition with unlabeled probe is shown in lanes 8–12. Supershift analysis with anti-STAT5A antiserum (lanes 13–16) compared with a control antibody (lanes 17–20) is also shown.