Fig. 3

Effects of cisplatin exposure on gene expression epimutations. A Boxplot of the number of new RNA epimutations arising at each generation of the MA lines compared to the pre-mutation generation F0 and for each condition: control (blue), cisplatin low dose (green) and cisplatin high dose (red). Each dot represents a generation in a lineage, and two lineages were used for each condition with control: N = 19, low dose: N = 20 and high dose: N = 19. No significant difference was observed between the conditions (Kruskal–Wallis rank-sum test, p-value = 0.11). B Survival curves representing the new RNA epimutations duration in each exposure condition: control (blue), cisplatin low dose (green) and cisplatin high dose (red). Curves were computed using the two lineages per condition as biological replicates. We observed a significant difference between the three conditions with an increased duration of epimutations in high dose and a decreased duration in low dose compared to control (log-rank test, p-value < 1e − 04). C Barplot of the mean percentage of new RNA epimutations that lasted more than one generation compared to the total epimutations arising each generation. Data are presented by condition: control (blue), cisplatin low dose (green) and cisplatin high dose (red). The means were calculated per generation from generation 2 to 18 with N = 9 for each condition. The mean percentages of lasting epimutations compared to all epimutations were 26.9% for control, 20.4% for low dose and 26.2% for high dose. No significant difference was observed between conditions. D Bubble plot illustrating ontology term enrichment of RNA epimutations lasting more than one generation in control (blue), cisplatin low dose (green) and cisplatin high dose (red) compared to gene without epimutation. Enrichments were calculated using data from generation 2 to 18 used as technical replicates, and two lineages per condition were used as biological replicates. Enrichment was calculated with χ.² test. The top 10 results per condition are shown. The x-axis shows the log10(χ) for enrichment. The y-axis shows the ontology terms. All displayed ontology terms were significantly enriched. E Bubble plot showing the distribution of lasting RNA epimutations in control (blue), cisplatin low dose (green) and cisplatin high dose (red). Data from generation 2 to 18 were used as technical replicates, and two lineages per condition were used as biological replicates. The y-axis displays the constitutive chromatin domains investigated. Active chromatin domains correspond to domains enriched in H3K36me3 mark and regulated domains to domains enriched in H3K27me3[44]. The x-axis shows the log2(odds) of enrichment. Odds ratio and p-values were calculated using Fisher’s exact test with Bonferroni correction. p-value cut-off for significance is 0.05. Supporting data are provided in the Excel file: “Additional file 14”