Fig. 1
BGA002 in combination with RA cooperates to inhibit MYCN activity in neuroblastoma. A-B cell lines are listed in the middle and grouped according to MYCN amplification status and/or p53 mutation status (MNA cell-lines: KELLY, CHP-134, IMR-32, SMS-KAN, LS, MHH-NB-11, NGP, SIMa, GI-LI-N; MNA-p53mut: SK-N-B [2] c, NMB, LAN-1; single copy: NBL-S, LAN-6, SH-SY5Y; single copy and p53mut: SK-N-F1). A a heatmap representing in vitro efficacy for mRNA inhibition after 12-hour treatment at 2.5 μM. The color scale represents the Log2 fold-change of MYCN mRNA with respect to untreated cells (n = 3 experiments for each cell line). B a heatmap representing in vitro efficacy for cell viability inhibition after 72 hours of treatment at different doses. The color scale represents the percentage of inhibition normalized over the control (n = 3 experiments). C decrement in viability by log10 GI50 and grouped by MYCN amplification and/or p53 mutation status. In the box plot, the median is indicated as the middle line, the outer edges of the box represent the first and third quartiles, and whiskers represent samples within the 1.5 interquartile ranges. Wilcoxon matched-pair test. D-E representative Western blot analysis after 48 hours of treatment (n = 3 experiments for each cell line). Left, representative staining for N-Myc (top) and associated GAPDH staining (bottom). N-Myc quantification normalized over the GAPDH is presented on the right. The bars represent the mean of 3 experiments, and the whiskers represent the standard deviation. D Western blot analysis for the Kelly cell line (MYCN amplified, p53 wild type). E Western blot analysis for the LAN-5 cell line (MYCN amplified, p53 wild type). F apoptosis measurement after 48 hours of treatment for the Kelly (left) and LAN-5 (right) cell lines (n = 3 experiments for each cell line). Bar plots represent the percentage of cells stained by AnnexinV+/PI+. The bars represent the mean, and the whiskers are the standard deviation. G transmission electromicrographs of cell lines treated for 48 hours. Kelly (top) and LAN-5 (down) representative images for each condition are presented (n = 2 experiments for each cell line). CTRL, control. Where indicated in the figure: *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001