Figure 3: Dabuzalgron Augments Mitochondrial Transcript Expression and Function in Hearts From Mice Treated With DOX

Male mice were treated with either DOX 20 mg/kg or vehicle by IP injection followed by 7 days gavage with either dabuzalgron 10 μg/kg twice daily, water, or trametinib (Trm) (1 mg/kg daily). Heart tissue was collected and immediately flash frozen on Day 7. (A) RNAseq was performed using RNA from the hearts of 3 mice per group (PBS + water; PBS + dabuzalgron; DOX + water; DOX + dabuzalgron). Gene set analysis was performed on DESeq2-derived statistics across these four categories. The results were highly enriched in gene sets involved in mitochondrial processes, a selection of which is shown here. (B) RNA abundance for all sequenced cytochrome C oxidase subunits (25 genes), mitochondrial complex I subunits (42 genes), and ATP synthase subunits (17 genes) was normalized by individual gene to vehicle treatment, then aggregated by treatment group. (C) Quantitative reverse transcription polymerase chain reaction for peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) was performed on mouse heart tissue (n in individual bars) (D) ATP content was measured in freshly harvested mouse heart tissue (total n in individual bars), then quantified relative to protein content. Results are presented relative to vehicle treatment for 4 independent experiments. (E) Thiobarbituric acid reactive substances (TBARS) were assayed in mouse myocardium. Abbreviations as in Figures 1 and 2.