Confocal immunofluorescent analysis of HeLa cells, serum starved (left), treated with hTGF-β3 #8425 (100 ng/ml, 30 min, center), or treated with hTGF-β3 and SB43152 (10 ug/mL, 1 hr, right), using Smad2/3 (D7G7) XP ® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

Flow cytometric analysis of HeLa cells using Smad2/3 (D7G7) XP ® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900 (red).

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and 10 μl of Smad2/3 (D7G7) XP ® Rabbit mAb, using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA using NEBNext ® Ultra™ II DNA Library Prep Kit for Illumina ® , and sequenced on the Illumina NextSeq. The figure shows binding across ID1, a known target gene of Smad2/3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and either 10 μl of Smad2/3 (D7G7) XP ® 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 SimpleChIP ® Human CDKN1A Intron 1 Primers #4669, SimpleChIP ® Human ID1 Promoter Primers #5139, 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.