Fig. 3

Highly specific, enhancer-based gene expression systems can be designed for other fusion-driven pediatric sarcomas. a Luciferase reporter assays of indicated fusion-positive ARMS (RH4 and RH30) and control cell lines after co-transfection with a reporter plasmid containing the alk-SE upstream of the minimal promoter YB-TATA and a constitutively expressed Renilla luciferase-encoding plasmid. Dots indicate Firefly to Renilla luminescence ratios normalized to a reporter plasmid without the alk-SE. Horizontal bars indicate mean and whiskers standard deviation per group. b Luciferase reporter assays of the same cell lines as in Fig. 3a after co-transfection with a reporter plasmid containing either syn_alk, syn_alk_3 or syn_alk_5 upstream of the minimal promoter YB-TATA and a constitutively expressed Renilla luciferase-encoding plasmid. Dots indicate Firefly to Renilla luminescence ratios normalized to a reporter plasmid without the alk-SE. Horizontal bars indicate mean and whiskers standard deviation per group. c Resazurin-based cell viability assay of pLenti_syn_alk_5_LT_Puro- or pLenti_0_LT_Puro-transduced and selected P3F1-positive ARMS cell lines 72 h after GCV addition. Dots indicate relative fluorescence units normalized to vehicle control for 4 biologically independent experiments. Lines show dose-response curves with 95% confidence interval based on a three-parameter log-logistic regression model calculated for each cell line. d Resazurin-based cell viability assay of P3F1-negative control cell lines transduced as in Fig. 3c. Dots indicate relative fluorescence units normalized to vehicle control for 4 biologically independent experiments. Lines show dose-response curves with 95% confidence interval based on a three-parameter log-logistic regression model calculated for each cell line. P-values were determined by two-tailed Mann-Whitney test, ****: p ≤ 0.0001