Fig. 4: RORα activation inhibits cell survival via BMAL1 and constrains lipogenic gene expression.
From: Restoration of the molecular clock is tumor suppressive in neuroblastoma
a Cell viability (MTT assay) and apoptosis (caspase 3/7 assay) in MNA (LAN5, IMR32) and non-MNA (SH-SY5Y, SK-N-AS) cells following treatment with SR1078 for 24 h (apoptosis) and 72 h (cell viability). IC50 values are shown in the lower panel. Cell viability data are mean ± SD and are representative from n = 2 biological replicates (n = 4 technical replicates). MNA cell lines in red; non-MNA cell lines in black. Apoptosis data are mean ± SD (n = 3; two-way ANOVA with Tukey’s multiple comparisons test). b Cell proliferation of LAN5 RORα Tet-ON cells cultured with (RORα-ON) and without (RORα-OFF) DOX over 8 days. Data are mean ± SD (n = 3; ****p < 0.0001; two-tailed unpaired t-test). Cell cycle analysis of LAN5 RORα-OFF and LAN5 RORα-ON cells following 10 days of DOX (2 µg/ml) induction. Data are mean ± SD (n = 2). c Left panel: cell growth of LAN5 SiCTRL and LAN5 SiBMAL1 cells in the presence and absence of SR1078 (5 µM) for 4 days. Data are mean ± SD (n = 3; two-tailed unpaired t-test). Middle panel: Caspase 3/7 activation in LAN5 SiCTRL and LAN5 SiBMAL1 cells following SR1078 treatment (10 µM for 24 h). Data are mean ± SD (n = 3; two-tailed unpaired t-test). Right panel: cell growth of LAN5 RORα SiCTRL and LAN5 RORα SiBMAL1 cells treated with (+Tet, RORα-ON) and without (−Tet, RORα-OFF) DOX for 4 days. Data are mean ± SD (n = 3, ****p < 0.0001; two-tailed unpaired t-test). d Left panel: Heat map of differentially expressed genes in LAN5 cells following SR1078 treatment (10 μM for 8 h). Right panel: Most enriched up- and downregulated pathways by SR1078 in LAN5 cells (p < 0.05). Blue = downregulated genes, red = upregulated genes.
