Western blot analysis of extracts from HeLa and C2C12 cells, untreated (-) or treated (+) with Trichostatin A (TSA) #9950 (1 μM, 18 hr), using Acetyl-Histone H2B (Lys15) (D8H1) XP ® Rabbit mAb (upper) and Histone H2B (V119) Antibody #8135 (lower).
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with TSA #9950 (right) using Acetyl-Histone H2B (Lys15) (D8H1) XP ® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5 ® #4084 (fluorescent DNA dye).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HeLa cells and either 10 µl of Acetyl-Histone H2B (Lys15) (D8H1) 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 GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP ® Human MyoD1 Exon 1 Primers #4490, 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.