Extended Data Fig. 7: LPS-driven mitochondrial stress triggers mitohormesis.
From: Mitohormesis reprogrammes macrophage metabolism to enforce tolerance
a. GO analysis of 1622 genes upregulated by both 4-OHE1 and LPS at 6h in BMDMs. b. Heatmap showing relative expression* of select genes upregulated by both 4-OHE1 and LPS at 6h in BMDMs. (*DESeq2 counts centered on the mean of each gene). c. RAW matrix-oxGFP macrophages pretreated with vehicle or 10 μM KRIBB11 for 1h before treatment with 4-OHE1 (5 μM, left) or LPS (right) for 8h. Matrix-oxGFP fluorescence was quantified by flow cytometry. d. MitoTracker Green signal in RAW macrophages and BMDMs measured by flow cytometry after 24h LPS simulation. e. Mitochondrial DNA/genomic DNA (mtDNA/gDNA) ratio in RAW macrophages treated with PBS vehicle control or LPS for indicated times. f. THP-1 and U937 matrix-oxGFP reporter cells treated 24h with LPS before matrix-oxGFP fluorescence was quantified by flow cytometry. g. WT (left) and Nrf2 KO (right) BMDMs were treated with EtOH vehicle control, or 4-OHE1/LPS (5 μM/100ng/mL) for 7h before Prdx6 qPCR. h. THP-1 cells were left untreated, or stimulated with LPS for 7h, before H2O2 treatment (500 μM, 5min) and intracellular oxidative stress measurement via CM-H2DCFDA staining and flow cytometry. Each data point is an independent biological replicate. For n = 2, data represented as mean; for n = 3, mean ± SEM. P values from unpaired, two-sided Student’s T Test (planned comparisons). RNA-seq study was performed once. Flow cytometry and qPCR data representative of 2 independent experiments.
