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. 2023 Jan 10;11(1):e005871. doi: 10.1136/jitc-2022-005871

Figure 4.

Figure 4

CI-994 treatment induces the expression of NF-κB pathway genes in MYC-driven medulloblastoma. (A–C) MED8A, D425 MED, and D341 MED treated with CI-994 or vehicle control were analyzed by RNA sequencing. The non-parametric supervised analysis identified 130 upregulated and 43 downregulated genes with a fold change ±2 and p≤0.05, which were analyzed by ingenuity pathway analysis (IPA). (A) Table showing top 20 out of 44 canonical pathways with activation z-score ≥2 and ranked by p value. (B) Table of top 10 upstream regulators identified by IPA with an activation score ≥1.8 and p≤0.05. (C/D) Unsupervised hierarchical clustering of genes regulated by NF-κB complex. (E) MED8A and D425 MED cells were treated with 2.5, 5, or 7.5 µM of CI-994 for 48 hours. TGM2 mRNA expression values are normalized to housekeeping controls, and expression is calculated relative to DMSO control. The values shown represent the mean±SD of three replicates per condition. (F) Western blots for TGM2 and Actin loading controls following CI-994 treatment for 24 and 48 hours. *, p<0.05; **, p<0.01; ***, p<0.001. DMSO, dimethyl sulfoxide; mRNA, messenger RNA; NF-kB, nuclear factor-kB; TGM2, transglutaminase 2.