Figure 4.

Stiffness-dependent oxidative phosphorylation modulates Treg induction. (A) Effect of OXPHOS inhibition on Treg induction. Oligomycin was added to the culture media at 0 hour, 24 hours, and 48 hours after seeding. For the Vehicle control, DMSO was added in the culture at the 0 hour time point (3 individual experiments and 2 replicates each, non-parametric, Kruskal-Wallis test with Dunn’s multiple comparisons was used for comparison between gels for each time point/vehicle, * p < 0.05, *** p < 0.001). (B) Mitochondrial potential on PA-gels. 72 hours after induction, cells were treated with MitoTracker Red, and mitochondria potential high is measured relative to the stained unstimulated control (7 individual experiments and 2 replicates each with 4 donors, parametric, unpaired t-test, **** p < 0.0001). (C, D) Effect of OXPHOS on mechanosensing of Treg induction. 250 μM of AICAR were introduced in the culture at 18 hours of the induction (7 individual experiments and 2 replicates each with 4 donors, one outlier found with the ROUT method in D was excluded, non-parametric, Mann-Whitney test was used to compare untreated to AICAR for each substrate, * p < 0.05).