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. 2024 Jan 27;15:830. doi: 10.1038/s41467-024-45044-1

Fig. 6. UA induces mitochondrial biogenesis and improves mitochondrial function.

Fig. 6

a Analysis of mitochondrial mass (GFP+ volume) in the retina of young or old mito-QC mice treated with UA or vehicle as shown in Fig. 4a, b (n = 5–7 mice). b Experimental design and readouts for the in vitro model of mtDNA release using the ARPE-19 cell line and the Bcl-2 inhibitor ABT-737 (10 μM). Pan-caspase inhibitor Q-VD-OPh (QVD, 10 μM) was added to inhibit apoptosis. Created with BioRender.com. c Representative images showing immunostaining of ARPE-19 cells for DNA (green) and TOMM20 (magenta, mitochondria), and corresponding quantification. UA (100 μM) was used as a mitophagy inducer and human cGAS inhibitor G140 (10 μM) was added as a control. (n = 3 independent experiments) d Western blot analysis of cGAS/STING mediators (TBK1, IRF3, cGAS, STING) in ARPE-19 cells (n = representative of 3 independent experiments). Protein levels are normalized to the loading control (Vinculin). e Representative images of mitophagy analysis in ARPE-19 cells using mito-QC reporter and corresponding analysis (n = 3 independent experiments). f Analysis of mitochondrial mass (MitoTracker Green; MTG) and mitochondrial superoxide production (MitoSOX Red/MTG ratio) by flow cytometry in ARPE-19 cells (n = 5 independent experiments). g Mitochondrial respirometry analysis in ARPE-19 cells using Seahorse XFe24 after sequential injection of Oligomycin, FCCP and Rotenone+Antimycin (n = 7–8 biological replicates from two independent assays). Oxygen consumption rate was normalized to cell number and graphs show basal respiration, ATP-linked respiration and spare respiratory capacity (SRC). Scale bars, 25 μm. All data are expressed as the mean ± s.e.m. Dots represent individual mice (a), independent experiments (c, e, f) or biological replicates from two independent assays (g). P values were calculated using 2-way ANOVA (a) or 1-way ANOVA with Šídák’s (c, e) or Tukey’s (f, g) post-hoc test (c, e). Source data are provided as a Source Data file.