Fig. 7.

Tonabersat enhances the upregulation of temozolomide‐induced genes involved in the regulation of DNA damage, stress response and apoptosis. (A) Graphical abstract for 3′‐mRNA sequencing performed with G35 cells (n = 3, technical triplicate). (B) Venn diagram depicting number of differentially expressed genes under indicated treatment conditions (P‐adj < 0.05, log2FC > 0.5). (C) GSEA for upregulated pathways under TMZ + TO vs. TMZ treatment. (D) Volcano plot showing the DEGs for TMZ treatment (left graph in box with dashed red line: TMZ vs. Ctr, Log2FC > 0.5, P‐adj < 0.05; right graph in box with red line: TMZ vs. Ctr, Log2FC > 2, P‐adj < 0.05). Wald test was used for hypothesis testing with Benjamini‐Hochberg for multiple correction testing. (E) Volcano plot showing the DEGs for TMZ + TO vs. Ctr (Log2FC > 2, P‐adj < 0.05). Combined treatment with TMZ + TO shows almost the identical profile. (F) Boxplots depicting the relative expression of DDIT3, ATF3 and GADD45A as representative genes for the regulation of DNA damage, stress response and cell cycle arrest. (G) Boxplot depicting the relative expression of PIF1. A pairwise t‐test was conducted to assess significance levels in “F, G,” based on three samples per treatment. The box represents the median ± IQR. Ctr, control; DEGs, differentially expressed genes; GSEA, gene set enrichment analysis; h, hours; Log2FC, log2 Fold Change; P‐adj, P‐adjusted; TMZ, temozolomide; TO, Tonabersat.