(A) Exercise protocol applied in this study. The mice were subjected to a single bout of acute treadmill running following a 4-day acclimatization protocol.

(B) Experimental design for the acute exercise at the early rest phase (ZT3) and the early active phase (ZT15) and tissue isolation every 4 h after sham exercise or exercise.

(C) Venn diagram displaying the number (top) and ratio (bottom) of oscillating genes in skeletal muscle isolated from sedentary (Sed) mice and exercised (Exe) mice at the early rest phase (ZT3).

(D) Heatmaps displaying rhythmic transcripts unique to Sed (left panel) and Exe (right panel).

(E) Venn diagram displaying cyclic genes from Sed and Exe mice at the early active phase (ZT15).

(F) Heatmaps displaying rhythmic genes found exclusively in Sed (left panel) and Exe (right panel).

(G) Amplitude distribution of oscillating genes in Sed and Exe mice at rest and active phase, respectively.

(H and I) Radar plots representing the phase lag of rhythmic genes unique to Sed (left panel) and Exe (right panel) mice at rest phase (H) and active phase (I).

(J and K) GO analysis of cyclic genes unique to Sed (left) and Exe (right) at rest phase (J) and active phase (K). TCR, T-cell receptor; PDGF, platelet-derived growth factor; ECM, extracellular matrix; TSR, thrombospondin type-1 repeat; TNF-α, tumor necrosis factor α; HIF1α, hypoxia-inducible factor 1α; TGF-β, transforming growth factor β; p75(NTR), p75 neurotrophin receptor; and F2,6BP, fructose 2,6-bisphosphate.

(L) Donut plots exhibiting the overlap of transcripts that oscillate only in Sed (transcripts lost oscillation), only in Exe (transcripts gained oscillation), both in Sed and Exe (rhythmic transcripts), and neither in Sed and Exe (non-rhythmic transcripts) between rest and active phase.

The transcriptomic data contain 5–6 individual gastrocnemius muscles per time point. The data were placed from ZT0 to ZT20 in the heatmaps and radar plots. Triangles indicate the timing of sham exercise or exercise. Nonparametric test Bio_Cycle was applied for the detection of the significance of daily oscillation using p value cutoff of 0.05.