Fig. 6

ATRA and 13-cis-RA downregulate expressions of MYCN and PLAGL2 in BE(2)-C cells. a-d Time-dependent effect of ATRA (a-b) and 13-cis-RA (c-d) on MYCN, PLAGL2 and miR-506-3p expressions. Cells were treated with RAs (5 μM) for up to 4 days. Shown are the RNA (a-c) and protein (b, d) levels of the indicated molecules. The data were analyzed as above. e-h Dose-dependent effect of ATRA (e-f) and 13-cis-RA treatments (g-h) on MYCN, PLAGL2 and miR-506-3p expression. Cells were treated with RAs at different concentrations for 4 days. The RNA (e, g) and protein (f, h) levels were detected as above. i-j CHIP-PCR assays under the treatment with ATRA. Cells were treated with ATRA (5 μM) or carrier DMSO (Control) for 4 days, and CHIP-PCR was performed as above. Protein levels in the Western blot images were quantified as above. i, PLAGL2 protein binding to its DNA-binding site in the promoter region of MYCN; j, N-Myc protein binding to its DNA-binding site in the promoter region of PLAGL2. Shown are the representative PCR results and quantifications