Fig. 5: Muscles from Icos^-/- NOD mice exhibit enhanced ROS production and transcriptomic oxidative stress features.

a Electron paramagnetic resonance (EPR) measurement of free radical (reactive oxygen species, ROS) production in the presence of ADP in muscle homogenates from Icos^-/- NOD and Icos^+/+ NOD mice at different time points corresponding to predisease, onset and established disease (for all Icos^+/+ NOD mice n = 6, for Icos^-/- NOD mice: n = 6 for 8 weeks, n = 7 for 25 weeks and n = 10 for 35 weeks). b Ex vivo analysis of H₂O₂ production in Icos^-/- NOD (n = 5) and Icos^+/+ NOD mice (n = 6) skinned muscles fibers in the presence of ADP. c RT-qPCR analysis targeted to oxidative stress-related genes of Icos^-/- NOD (n = 8 for 8 and 25 weeks, n = 11 for 35 weeks) vs. Icos^+/+ NOD (n = 8/group for 8 and 35 weeks and n = 6 for 25 weeks) mice (arb. units: arbitrary units). For a and b, mean values ± s.e.m are shown. For c, each box expands from max to min values and median are represented. For a and b, mean ± s.e.m and a representative experiment out of two are shown. For a, statistical analyses were performed using the Two-way ANOVA test with Sidak’s post-hoc multicomparison. For b, statistical analyses were performed using the Mann–Whitney test (two-tailed) (left) and correlations by Pearson’s test. For c, statistical analyses were performed using the Kruskal-Wallis test with uncorrected Dunn’s test (for each genotype, comparison shown in the graph corresponds to comparison with values from mice at 8 weeks). P values displayed correspond to the comparison of 25 and 35-week-old mice data of one given genotype with 8-week-old mice data of the same genotype. For all panels, n correspond to the number of independent mice. Numbers on panels denote p values. Source data are provided as a Source Data file.