Figure 10. SLFN12 increases the cytotoxic effect of carboplatin in Hs578t and BT549 cells. (A, B) Primer-probe RT-qPCR analysis of SLFN12 mRNA levels, 48-h after transduction with adenoviral vector expressing SLFN12 (AdSLFN12) or control empty vector (AdCMV) in Hs578t (A) and BT549 (B) cells. HPRT was used as a reference gene. (n=3, *** p≤0.001, **** p≤0.0001). (C, D) Number of Hs588T (C) and BT549 (D) cells, measured by crystal violet uptake, after 72 h of transduction with viral vector expressing SLFN12 (AdSLFN12) or the control vector (AdCMV) in combination with 43 μM of carboplatin (added simultaneously with the viral vector transduction). AdSLFN12 transduction reduces cell numbers, while the simultaneous addition of carboplatin further reduces cell number (data normalized to AdCMV group, n=15, **p<0.01, ***p≤0.001, ****p≤0.0001, one-way ANOVA). (E, F) Cell cytotoxicity calculated from the cell number data shown in Figures C and D by measuring the ratio of each carboplatin-treated group to its own control without carboplatin shows that the combination of carboplatin (43 μM) and SLFN12 over-expression (AdSLFN12) increases the cytotoxic effect of carboplatin in (E) Hs578t, and (F) BT549 cells. (***p≤0.001, ****p≤0.0001, Student’s t-test).