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. 2010 Aug 4;9:208. doi: 10.1186/1476-4598-9-208

Figure 5.

Figure 5

Roscovitine when combined with Doxorubicin increases DNA DSBs and overall DNA damage over time in A549 cells. A) Western blot analysis of DNA DSBs by phosphorylated γH2AX (serine 139) immediately (t0) or 6 (t6) and 24 (t24) hours following a 6 hour treatment with either 750 nM Doxorubicin (D) or 20 μM Roscovitine alone or in combination (DR). B) Immunofluorescence analysis by fluorescent microscopy of phosphorylated γH2AX (serine 139) at the abovementioned time points following 6 hours of treatment with 20 μM Roscovitine or 2.5 μM Doxorubicin (as a positive control for DSBs). Images shown are γH2AX (FITC) and DAPI merges under 100× (upper panels) and 400× (lower panels) magnifications. C) Alkaline comet assay quantification and D) respective images (400x magnification), 6 (t6) and 24 (t24) hours following a 6 hour incubation with abovementioned treatments (Control, NT; Doxorubicin, D; Doxorubicin + Roscovitine, D+R; Roscovitine, R) to measure overall DNA damage.