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. 2020 Dec 23;13(1):21. doi: 10.3390/cancers13010021

Figure 5.

Figure 5

5-azaC induced DNA double strand breaks in HPV+ HNSCC depend on active transcription. (A) Schema of 72 h 5-azaC treatment with transcriptional inhibition during the last 24 h before analysis. (B) PFGE depicting HPV+ UMSCC47 cells after 72 h of 20 µM 5-azaC and addition of 1 µM triptolide, 1.25 µg/mL actinomycin D, or 10 µM DRB for the last 24 h. (C and D) Coimmunostaining of EU (active transcription) and ɣH2AX in HPV-positive UMCC47 cells treated or not with 10 µM of 5-azaC in the presence or absence of RNA polymerase I inhibitor elipticine. Scale bars are 50 (C) and 10 (D) µm. (E) Clonogenic survival of UMSCC47 cells either treated or not with 1 ng/mL actinomycin D 48 h after initiation of increasing doses of 5-azaC treatment (log scale). Standard deviations calculated from two independent experiments. Student’s t test was performed to test significance. * indicates p-value < 0.05.