Fig. 6.

Assays to detect DNA damage and apoptosis in day 13 PHK and HPV-18−infected raft cultures. (A) Detection of cleaved caspase 3 (green) and γ-H2AX (S139) (red) in HPV-18 raft cultures following exposure to DMSO or Vorinostat (1 and 5 μM) from days 6 to 13. (B) HPV-18 raft cultures were probed for TUNEL (green) (Upper), whereas PHK raft cultures were probed for TUNEL (green) and γ-H2AX (red) (Lower). (C) IB detection of γ-H2AX S139 in parallel HPV-18 raft cultures. Lysates from UT PHK or infected raft culture as well as infected culture exposed to vehicle (0) were used as references. Actin served as a loading control. (D) Percentage of TUNEL-positive nuclei in A and B were averaged from four microscopic fields under a 20× objective. (E) IB detection of cleaved caspase 3 in parallel Vorinostat-treated HPV-18 raft and untreated PHK raft cultures. This strip of IB was derived from the same gel as depicted in Fig. 5E, and the actin loading control was presented in Fig. 5E.