Fig. 3.

Detection of acetylated histone 4 in the presence or absence of Vorinostat. (A) Acetylated histone 4 was detected with antibody reactive to H4 K5/6/9/12/16 (red signals) in IBs of lysates from two independent sets of day 13 HPV-18−infected cultures (a and b). The cultures were exposed to DMSO (0) or to 0.2, 1, or 5 μM Vorinostat from days 6 to 13. Acetylated H4 (n red) and actin (in green) were recorded with the LI-COR CLx system. (B) Acetylated H4K12 was detected in the IB of the above lysates using the enhanced chemiluminescence (ECL) method. Lysate from UT PHK was one of the controls. This IB (marked with an asterisk) was derived from the same gel as depicted for HDAC-2 in Fig. 4A and for HDAC-5 and actin loading reference in Fig. 4B. Indirect IF detection of elevated (C) H4K12Ac and (D) H4K16Ac in day 13 HPV-18 raft cultures in the DMSO-treated control (Upper) or exposed to 5 μM Vorinostat (Lower) from days 6 to 13. (E) Indirect IF detection of H4K12Ac (green) in day 13 HPV-18 raft cultures treated with 2 μM Vorinostat from days 6 to 13. In C–E, merged images are shown in which nuclei were detected with DAPI and acetylated H4K in green (Left), while only the acetylated H4K signals were shown (Right).