Fig. 3.
Mechanism of synergistic effect of the dual-drug combination. a Heatmap shows the H3K27me3 pattern around the transcription start site (TSS) region. Each panel represents 0.5 kb upstream and downstream of the TSS. b Heatmap of H3K4me3 and H3K27ac ChIP-seq signals in cells treated with DMSO and J4 at promoter-proximal regions. Rows are sorted by exclusive occupancy in the J4 condition. Cluster 1 in the metaplots refers to the occupancy particular in the DMSO condition, and cluster 2 represents the inverse comparison. c Venn diagram showing the overlap of upregulated genes by J4 among H3K4me3 and H3K27ac ChIP-seq peak. d GO analysis of 2092 commonly shared genes from H3K4me3 and H3K27ac ChIP-seq. e Genome browser tracks of H3K27me3, H3K4me3, and H3K27ac occupancy and RNA-seq at the CHOP gene locus. f RNA-seq analysis of differentially expressed genes in SJSA-1 cells treated with different drug formulations. g Venn diagrams showed the overlaps of differential genes of the three groups compared to the control. h Gene Set Enrichment Analysis (GSEA) showed that ER stress-associated genes were enriched in dual-drug-specific differential genes. i Fold change in ER stress-associated genes in SJSA-1 cells treated with different formulations for 48 h by qPCR analysis. j Western blot analysis of CHOP, GADD34, and ATF4 expression after the treatment with free dual drug or NPJ4+Apa (J4 5 μM; Apa 10 μM) for 48 h. β-Actin was used as the internal control. ***P < 0.001, **P < 0.01, or *P < 0.05