Qscript cdna supermix 5

5 × 10⁴ Res186 and Res199 WT or drug resistant cells were grown in 6-well plates (BD Falcon) and treated following the protocol described for quantification of nuclear fragmentation. Total RNA was isolated with the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). cDNA was synthesized with qScript cDNA SuperMix (5 ×) (Quanta Biosciences, 95048-025) following real-time (RT) PCR with human ABCB1 forward 5′-cccatcattgcaatagcagg-3′ and reverse 5′-gttcaaacttctgctcctga-3′ primers and GAPDH forward 5′-tgacatcaagaaggtga-3′ and reverse 5′-tccaccaccctgttgctgta-3′ primers as described previously [13 (link)]. To ensure accuracy, an internal reference reaction was performed on the same sample as used for the target gene. The results were standardized with the formula: ΔCT = CT_Ref − CT_Target and converted to folds of target gene over reference gene (F = 2^{- ΔCT}). Data from a minimum of 3 independent experiments was used to quantify gene expression. P values of less than 0.05 were considered statistically significant.