MYCN directly binds to the promoter regions of NeuroD1. (A) Schematic
describing the MYCN responsive E-box (CACGTG) motif within the NeuroD1 gene
promoter. TSS represents the transcription start site. (B, C) Chromatin
immunoprecipitation (ChIP) assays were performed by using anti-MYCN antibodies
in IMR-32 and CHP-134 cells, and an IgG antibody was used as the control. PCR
was then performed to examine the bound DNA fragments. The results represent
the mean ± SD (n = 3).
**p < 0.001 versus the IgG group. (D)
Human embryonic kidney (HEK) 293T cells were cotransfected with pRL-CMV
(Renilla) control reporter vector, pGL4.74-NeuroD1 or
pGL4.74-mut construct, MYCN expression vector, or control-expressing vector.
The luciferase activity was measured and normalized by Renilla
luminescence signals. The results represent the mean ± SD
(n = 3).
**p < 0.001 versus the control-expressing
vector plus pGL4.74-NeuroD1-expressing group;
#p < 0.01 versus the CMV-MYCN plus
pGL4.74-NeuroD1-expressing group.