Figure 5. THZ1 Inhibits MYCN-driven Transcriptional Amplification.
(A) Box plot depicting the log2 fold changes in actively transcribed genes in MYCN-nonamplified SH-SY5Y cells transduced with MYCN (pLenti 6.3-MYCN) compared with a control vector (pLenti 6.3). Box plot whiskers extend to 1.5 times the interquartile range (n=18,665 expressed genes; p<2.2 × 10−16, two-sided Mann-Whitney U test).
(B) Heat map of all gene expression values in MYCN-nonamplified SH-SY5Y cells vs. SH-SY5Y cells overexpressing MYCN (pLenti 6.3-MYCN) following either THZ1 (100 nM for 6 hr) or DMSO treatment. Rows show z-scores calculated for each vector.
(C) Immunoblot analysis of MYCN, CC3 and cleaved PARP in SH-SY5Y MYCN-nonamplified cells engineered to express either a control vector (pLenti 6.3) or MYCN (pLenti 6.3-MYCN) following treatment with THZ1.
(D) Heat map of differentially expressed genes in MYCN-amplified NB cells expressing a MYCN shRNA vs. control shRNA. Rows show z-scores calculated for each vector.
(E) Box plots of log2 fold changes in gene expression in MYCN-amplified NB cells expressing a MYCN shRNA vs. a control shRNA. Box plot whiskers extend to 1.5 times the interquartile range (n=18,665 expressed genes; p<10−15 for shMYCN vs. shGFP, two-sided Mann-Whitney U test).
(F) GSEA plot depicting the correlation between the top 500 downregulated genes following THZ1 treatment and the rank-ordered genes that are differentially expressed after MYCN knockdown in MYCN-amplified NB cells.
See also Figure S5.