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. Author manuscript; available in PMC: 2020 Jan 14.
Published in final edited form as: J Immunol. 2010 Oct 1;185(9):5074–5081. doi: 10.4049/jimmunol.1001725

FIGURE 5.

FIGURE 5.

Mutation within the STAT5 binding site abolishes its ability to bind to STAT5. EMSA with the putative STAT5 binding site sequence of the granulysin enhancer was performed. 32P-labeled STAT5 consensus (C) probe (lane 6) and STAT5 wild-type (Wt) probes based on granulysin enhancer sequence (lanes 1–5), were incubated with nuclear extracts from CRL-2105 T cells stimulated with IL-2 at concentration of 100 U/ml or 400 U/ml as indicated. For competition assays, a 100-fold molar excess of unlabeled STAT5 Wt (lane 4), STAT5 mutant (Mut; lane 5) oligonucleotides were incubated with CRL-2105 nuclear extracts prior to incubation with 32P-labeled STAT5 Wt probe. Lane 7, Labeled STAT5 Wt probe without nuclear extract. The labeled STAT5 Wt probe incubated with CRL-2105 nuclear extracts after incubating with 400 U/ml IL-2 (lane 2) or 100 U/ml IL-2 (lane 3). Ab-mediated supershift assays were performed with 2 mg rabbit anti-STAT5 Ab and NRS as a control.