Western blot analysis of extracts from various cell lines, untreated (-) or treated with either DMOG (1 mM, 6 hr; +) or cobalt chloride (100 µM, 4 hr; +) as indicated, using HIF-1α (D2U3T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 MCF7 cells treated with cobalt chloride (100 μM) overnight and 10 μl of HIF-1α (D2U3T) Rabbit mAb, using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext ® Ultra™ II DNA Library Prep Kit for Illumina ® , and sequenced on the Illumina NextSeq. The figure shows binding across the PCAT6 gene. For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunopreciptations were performed with cross-linked chromatin from 4 x 10 6 MCF7 cells treated with cobalt chloride (100 μM) overnight and either 10 μl of HIF-1α (D2U3T) 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 ARRDC3 Downstream Primers #75671, human ERRFI1 upstream primers, and SimpleChIP ® Human α Satelite 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.