(A) Analysis of event-free survival split by MYCN amplification status in NB with low (bottom 50%; gray) and high (top 50%; black) TGFBR3 expression in the Oberthuer data set (36). Amp, MYCN amplified (dashed lines); NA, nonamplified (solid lines). Numbers in parentheses indicate the number of samples. (B) Microarray data set analysis for TGFBR3 expression. Data are presented as median (horizontal bars) and interquartile range (boxes). ****P < 0.0001 (Mann-Whitney). (C) Linear regression of MYCN and TGFBR3 expression in the microarray data set. (D) Western blot and I¹²⁵ TGF-β binding and crosslinking with TβRIII pull-down of SK-N-AS-MYCNER–inducible cell line in the presence and absence of 4-hydroxytamoxifen (4OHT) to stabilize MYCN. (E) SHEP-21N–repressible cell line in the presence and absence of doxycycline (Dox) to repress MYCN expression. Dox was replenished at day 3 for the 5-day treatment in the binding experiment. (F) ChIP in SHEP-21N cells using primers for Sp-1 binding sites in TβRIII. Data are representative of 3 experimental replicates with similar trends. (G) I¹²⁵ TGF-β binding and crosslinking with TβRIII pull-down in the presence and absence of trichostatin A (TSA) (1- and 4-hour treatments) and valproic acid (VPA) (3- and 6-day treatments) at the concentrations shown. Western blots for acetyl-lysine (Ac Lys) and TβRIII in the presence and absence of trichostatin A (4-hour treatment). Background and β-actin–normalized integrated density for TβRIII are shown as percent control.