Western blot analysis of extracts from TF-1 cells, treated with GM-CSF (25 ng/ml) for the indicated times, using Phospho-Stat5 (Tyr694) Antibody (upper) or Stat5 antibody (lower).
Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue), using Phospho-Stat5 (Tyr694) Antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 10 μl of Phospho-Stat5 (Tyr694) Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP ® Mouse CIS Intron 1 Primers #5131, mouse SOCS-3 promoter primers, and SimpleChIP ® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.