Western blot analysis of extracts from HeLa, C2C12, and C6 cells, untreated (-) or treated with Trichostatin A (TSA) #9950 (1 μM, 18 hr; +), using Acetyl-Histone H4 (Lys16) (E2B8W) Rabbit mAb (upper) or Histone H4 (L64C1) Mouse mAb #2935 (lower).
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (1 μM, overnight; right) using Acetyl-Histone H4 (Lys16) (E2B8W) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A (TSA) #9950 (1 uM, Overnight; green) using Acetyl-Histone H4 (Lys16) (E2B8W) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab') 2 Fragment (Alexa Fluor ® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HeLa cells and either 10 μl of Acetyl-Histone H4 (Lys16) (E2B8W) 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, 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.