Western blot analysis of extracts from various cell lines using Stat3 (79D7) Rabbit mAb.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence ® Control siRNA (Unconjugated) #6568 (-), SignalSilence ® Stat3 siRNA I (+) or SignalSilence ® Stat3 siRNA II #6582 (+), using Stat3 (79D7) Rabbit mAb #4904 and α-Tubulin (11H10) Rabbit mAb #2125. The Stat3 (79D7) Rabbit mAb confirms silencing of Stat3 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Stat3 siRNA.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 Hep G2 cells treated with IL-6 (100 ng/ml) for 30 minutes, and either 20 μl of Stat3 (79D7) 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 c-Fos Promoter Primers #4663, 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.