Supplemental Files

Supplementary Files for manuscript.
Supplementary Table S1. List of abbreviations used in manuscript.
Supplementary Figure S1. Fiber characteristics of skeletal muscle biopsies from COCOA-PAD trial. A-C) Change in average myofiber cross-sectional area (CSA) of Type I (A), Type IIa (B), and Type IIa/x (C) fibers from baseline to 6-month follow-up in PLA and COCOA groups. D-F) Change in fiber type frequency, % of total fibers that are Type I (D), Type IIa (E), or Type IIa/x (F). G) Change in the proportion of Type I fibers (%) with central nuclei. Difference between groups tested using an independent t-test, n=5 in PLA, n=9 in COCOA, ns: not significant.
Supplementary Figure S2. Correlations between fiber characteristics, muscle measures, and walking performance from COCOA-PAD trial. A-B) Association between the fold change in HO-1 levels (A) or NQO1 levels (B) with the change in Type II fibers with central nuclei. (C-D) The association between the fold change in HO-1 levels (C) or NQO1 levels (D) with the fold change in UQCRC2 levels. E) Association between the fold change in UQCRC2 levels and the change in 6-minute walk distance (m) from baseline to 6-month follow-up. Association tested by Pearson correlation. COCOA points shown in green and PLA shown in gray. Line of best fit, R2, and p-values shown.
Supplementary Figure S3. PAD serum treatment and EPI treatment effects on myotubes. A) OCR trace over time after injections of indicated mitochondrial respiration modulators in Control myotubes [CON, standard 2% horse serum (HS)], 10% pooled PAD serum-treated myotubes (PAD serum), and 10% pooled healthy participant serum-treated myotubes (Healthy serum). B) Quantification of maximal OCR (FCCP-stimulated), normalized to protein concentration. C) Hydrogen peroxide production, assessed by AmR fluorescence. Data expressed as fold-regulation relative to CON myotubes. Experiments are averages of 4 technical replicates. D) Representative Western blot of p-Nrf2 and Nrf2 levels in myotube lysates treated with increasing doses of EPI or sulforaphane (SFN) as a positive control. E) Quantification of p-Nrf2 levels after densitometric analysis of the levels of each sample normalized to corresponding Nrf2 (p-Nrf2/Nrf2), expressed as fold-regulation relative to CON (untreated, 0 μM). Difference between groups (n=2-3/group) tested using a one-way ANOVA, results of pairwise post-hoc analyses (Tukey’s) displayed as *p<0.05, **p<0.01, ***p<0.001.