Schematic of study design. (A) Experimental design. Human vastus lateralis biopsies were collected before and 3.5 h after 40 min of endurance exercise at 70% VO2max in four participants. Biopsies were also collected at the same time points from two supine resting circadian controls. Nuclei were isolated and underwent same-cell single-nuclei multiome assay to assess gene expression and chromatin accessibility simultaneously. (B) Computational design. High-quality single-nuclei assay data were integrated and clustered. Single-cell bioinformatic analyses (identification of differentially expressed genes and differentially accessible regions, pathway analysis, and characterization of coordinated expression and chromatin changes using latent variable analysis) were performed for each cell type followed by both gene-centric and TF-centric regulatory circuit analyses. Regulatory circuit analyses were used to infer biological insight and compiled in a user-friendly web resource.
