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. 2024 Jul 11;15:5585. doi: 10.1038/s41467-024-49871-0

Fig. 4. MYCN drives increased RUNX1T1 translation.

Fig. 4

a Western blot of SH-EP TET-21/N cells after 72 h of treatment with 1 µg/mL doxycycline (left panel). Quantitation from three independent experiments demonstrated significantly decreased MYCN (****p = 0.0001) and RUNX1T1 expression (***p < 0.0001) (right panel). Values represent means from three independent experiments ±SD, two-tailed unpaired t-test. b MYCN and RUNX1T1 mRNA expression after 72 h doxycycline treatment, showing significant downregulation of MYCN (***p = 0.0009) but no significant change in RUNX1T1 (p = 0.9559). Values represent means from three independent experiments ±SD, two-tailed unpaired t-test. c Western blot of SH-EP neuroblastoma cells overexpressing MYCN and empty vector (EV) control (left panel). Quantitation of western blot, from three independent experiments (right panel) demonstrated significantly increased RUNX1T1 expression (*p = 0.0339). Values represent means from three independent experiments ±SD, two-tailed unpaired t-test. d RUNX1T1 and MYCN mRNA expression in SH-EP cells overexpressing MYCN, relative to the control gene (GUSB). Values represent means from three independent experiments ±SD. Two-tailed unpaired t-test, ****p < 0.0001; ns: not significant (p = 0.5679). e Cyclohexamide chase experiment showing RUNX1T1 protein stability in three neuroblastoma cell lines (BE(2)-C, KELLY, SH-EP) over a 12 h time course. Values represent means from three independent experiments ±SD. f Luciferase activity following transfection of murine Runx1t1 5’UTR into MYCN overexpressing or EV SH-EP cells. Values represent means from three independent experiments ±SD. Groups were analysed using two-way ANOVA and Bonferroni multiple comparison. ***p = 0.0007; ns not significant (p = 0.9335). Source data are provided as a Source Data file.