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Fig. 4 | Journal of Hematology & Oncology

Fig. 4

From: Preclinical anti-myeloma activity of EDO-S101, a new bendamustine-derived molecule with added HDACi activity, through potent DNA damage induction and impairment of DNA repair

Fig. 4

EDO-S101 reduces HR efficiency. a JJN3HR and U266HR cells were pretreated with EDO-S101 for 24 h, simultaneously transfected with 5 μg of I-SceI endonuclease-expressing plasmid and 0.5 μg of pDSRed2-N1 (red cells), and incubated in the presence or absence of EDO-S101 for 30 additional hours at indicated doses. Correct repair by HR of the DSB induced by the endonuclease restored a functional GFP gene whose expression could be detected as green cells. b Flow cytometry analyses of 100000 GFP+ and/or DsRed+ cells are shown for JJN3HR and of 200000 cells for U266HR. Efficiency of HR is showed on the right side and was calculated 30-h post-transfection as the ratio of GFP+ (green) to DsRed+ cells (red). Data are expressed as the mean of a minimum of three independent experiments ± SD (***p < 0.001, compared to untreated cells). c Immunofluorescence assay for γH2AX and RAD51 in JJN3HR cells after 5-h post-irradiation with 2 Gy with or without EDO-S101 treatment. Percentage of foci with double staining for γH2AX and RAD51 are shown on the right side. Data are the mean of three independent experiments. One hundred cells were counted in each experiment

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