Fig. 2

N-Myc and c-Myc upregulate JMJD6 expression by binding to the JMJD6 promoter. a, b CHP134 cells were transfected with control siRNA, N-Myc siRNA-1 or N-Myc siRNA-2, and SK-N-AS cells were transfected with control siRNA, c-Myc siRNA-1, or c-Myc siRNA-2. Forty-eight hours later, RNA and protein were extracted for RT-PCR (a) and immunoblot (b) analyses. Error bars represent normalized standard errors from three independent experiments (**p < 0.01, ***p < 0.001, one-way ANOVA). c SHEP Tet/21N cells were treated with DOX (2 µg/ml) or vehicle control for 48 h. RT-PCR and immunoblot analyses were conducted. Error bars represent normalized standard errors from three independent experiments (***p < 0.001, two-tailed unpaired Student’s t test). d Schematic representation of the JMJD6 gene promoter. TSS indicates transcription start site. Amplicons A, B, and C represented the sites for ChIP PCR primers. e ChIP assays were performed with a control IgG, anti-N-Myc or anti-c-Myc antibody (Ab), followed by PCR with primers targeting a remote negative control region (amplicon A), the E-box region (amplicon B), or the exon 2 region (amplicon C) of the JMJD6 gene in CHP134 and SK-N-AS cells. Fold enrichment of the JMJD6 gene region was calculated as the difference in cycle thresholds obtained with the anti-N-Myc or anti-c-Myc Ab compared with the control IgG, relative to input. Error bars represent normalized standard errors from three independent experiments (*p < 0.05, **p < 0.01, two-tailed unpaired Student’s t test). Source data are provided as a Source Data file