Western blot analysis of extracts from NK-92 cells, untreated or treated with Human Interleukin-2 (hIL-2) #8907 (2ng/ml, 15 minutes), using Phospho-Stat4 (Tyr693) Antibody (upper) or Stat4 (C46B10) Rabbit mAb #2653 (lower).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 NK-92 cells starved of IL-2 overnight then treated with IL-12 (10ng/ml) for 4h and either 20 μl of Phospho-Stat4 (Tyr693) Antibody #5267 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 human IRF-1 promoter primers, SimpleChIP ® Human PRF1 Promoter Primers #9014, and SimpleChIP ® Human α Satellite Repeat Primers #4486. 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.