Fig. 3

SMAD9 knockdown suppresses MYCN-amplified NB growth. A SMAD9 knockdown in MYCN-amp cells (BE2-C, IMR-32 and SK-N-BE2), as measured using Q-RT-PCR. B The viability of MYCN-amp cells was measured using CTG after knockdown of SMAD9. C, D Colony formation of MYCN-amp cells after SMAD9 knockdown (C) and quantification of CVS results (D). E, F, G, H SMAD9 knockdown in non-MYCN-amp cells (SK-N-SH and SH-SY5Y in E) measured using Q-RT-PCR (E) and cell viability (F) and colony formation (G, H) after SMAD9 knockdown. I, J Exogenous SMAD9 overexpression without (WT) or with (MUT) SMAD9-CDS mutation and shSMAD9 transfection. SMAD9 mRNA levels in MYCN-amp cells measured using Q-RT-PCR (I) and cell viability after SMAD9 overexpression (J). CDS: coding sequence; CTG: Cell-Titer-Glo; CVS: crystal violet staining; MYCN-amp: MYCN amplification; MUT: mutation; non-MYCN-amp: non-MYCN amplification; Q-RT-PCR: quantitative reverse transcription polymerase chain reaction; shRNA: short hairpin RNA; shSCR: shRNA scrambled control; WT: wild type. * P < 0.05, **P < 0.01 and *** P < 0.001