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. 2019 Nov 1;20(21):5455. doi: 10.3390/ijms20215455

Figure 2.

Figure 2

Effects of MK-8776 exposure on HPV-18 E6 and E7 and DNA damage response (DDR) proteins, as well as a biomarkers of DNA damage and apoptosis in raft culture lysates. (A–C) Uninfected (PHK) and infected (HPV-18) raft cultures were exposed to DMSO or 20 µM MK-8776 from days 7 to 12 and harvested on day 12. Lysates were probed by immunoblot assays to detect HPV-18 E6, E7, pRB, p130, p53, ph-Chk1 S345, ph-ATR S1986 and γ-H2AX S139. (D–H) Immunoblot analysis of lysates from HPV-18 raft cultures exposed to DMSO, 10 or 20 µM MK-8776 from days 7–12 or days 7–14 and harvested on days 12 or 14, respectively. Immunoblots were performed to detect total ATR, total ATM, ph-ATM S1981, total Chk2, ph-Chk2 T68, ph-Chk1 S317, total Chk1, ph-Chk1 S296 and cleaved caspase. Actin served as loading control for each set of immunoblots. Arrowhead points to p130 protein band. * denotes a non-specific protein detected by the p130 antibody.