Western blot analysis of extracts from NK-92 cells, untreated (-) or treated (+) with Human Interleukin-2 (hIL-2) #8907 (10 ng/ml, 15 min) or human interleukin-12 (hIL-12, 50 ng/ml, 15 min), using Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb (upper) and Stat4 (C46B10) Rabbit mAb #2653 (lower).
Confocal immunofluorescent analysis of NK-92 cells, starved of IL-2 (5 hr) and then either untreated (upper) or IL-12-treated (50 ng/mL, 15 min; lower), using Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).
Flow cytometric analysis of NK-92 cells, untreated (blue) or treated with IL-12 (green), using Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb.
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 (10 ng/ml, 4 hr) and either 20 μl of Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb 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.