Western blot analysis of extracts from various cell lines using c-Myc (D3N8F) Rabbit mAb.
Western blot analysis of extracts from HT-29 cells, untreated (-) or treated with JQ1 (1 μM; 72 hr; +), using c-Myc (D3N8F) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, the BET bromodomain inhibitor JQ1 inhibits c-Myc expression.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence ® Control siRNA (Unconjugated) #6568 (-) or SignalSilence ® c-Myc siRNA I #6341 (+), using c-Myc (D3N8F) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The c-Myc (D3N8F) Rabbit mAb confirms silencing of c-Myc expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
Confocal immunofluorescent analysis of HT-29 cells, untreated (left) or treated with bromodomain inhibitor JQ1 (1 μM, 72 hr; right), using c-Myc (D3N8F) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 Daudi cells and 10 μl of c-Myc (D3N8F) 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 NPM1, a known target gene of c-Myc (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 Daudi cells and either 10 μl of c-Myc (D3N8F) 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 ATF4 promoter primers, SimpleChIP ® Human NPM1 Intron 1 Primers #4779, 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.