Fig. 10. JTE-607 leads to DNA damage and S phase crisis.

a Representative flow cytometry data of γH2A.X expression levels in U937/iFIP1 cells in Dox− or Dox+ conditions after JTE-607 treatment for 24 h. b Fold change of γH2A.X signals in U937/iFIP1-A1 cells with or without FIP1 induction after JTE-607 or DMSO treatment. P value is based on the Chi-squared test. Fold change is based on JTE-607-treated vs. DMSO-treated samples in Dox− and Dox+ conditions, respectively. c Cell cycle analysis using U937/iFIP1-A1 cells. Data are based on 24 h post JTE-607 treatment. d Percent of cells in different cell cycle phases as based on data in c. e Ratio of cells in the 1st half S phase to the 2nd half S phase after JTE-607 treatment in U937/iFIP1 cells in Dox+ vs. Dox− conditions (n = 2). P value (t-test) for significance of difference is indicated. f Synergy scores for JTE-607 with another indicated compound in U937/iFIP1 cells in Dox+ or Dox− conditions. Synergy scores were calculated by the SynergyFinder program using the Bliss method. The ‘+/− value’ indicates 95% confidence interval (two replicates). The function of each compound is indicated, namely, TOPIi, TOPIIi, and ATRi. g Representative synergy score plots generated by the SynergyFinder program for JTE-607 in combination with BAY 1895344 in U937/iFIP1-A1 cells in Dox− (left) or Dox+ (right) conditions. h As in f, except that synergy analysis was carried out in U937 cells (two replicates). i A model summarizing mode of action of JTE-607 (CPAi). Alternative polyadenylation and transcriptional readthrough lead to transcriptomic disturbance, the extent of which correlates with the CPA activity of a cell. Transcriptional readthrough also leads to S phase crisis, due likely to transcription-replication conflict, and then DNA damage, which is exacerbated by a high cell proliferation rate. CPAi causes cell death through both transcriptomic disturbance and DNA damage. Source data are provided as a Source Data file.