FoxO maintains a genuine quiescent muscle stem-cell state until geriatric age (RNA-seq)

We identify two quiescent stem-cell states through relative CD34 expression: CD34High, with stemness properties (genuine state), and CD34Low, more committed to myogenic differentiation (primed state). The genuine-quiescent state is preserved into later life succumbing only in extreme old age due to acquisition of primed-state traits. We identified niche-derived IGF1-dependent Akt activation as detrimental to the genuine stem-cell state by imposing primed-state features via FoxO inhibition. Interventions to neutralize Akt and promote FoxO activity drive primed-to-genuine state conversion, while FoxO inactivation deteriorates the genuine state at young age, causing muscle regenerative failure, as in geriatric mice. Overall design: Muscles form mice were mechanically disaggregated and dissociated in Ham's F10 media containing Liberase (0.1mg/g muscle weight) (Roche; 5mg/mL) and Dispase 0.3% at 37?°C for 2 h and then filtered. Cells were then incubated in lysis buffer (BD Pharm Lyse) for 10?min on ice, re-suspended in PBS with 2% goat serum (FACS buffer) and counted. PE-Cy7-conjugated anti-CD45 (Biolegend 103114), anti-Sca-1 (Biolegend 108114) and PE-Cy7-conjugated anti-CD31 (Biolegend 102418) antibodies were used for lineage-negative selection. PE-conjugated anti-a7-integrin (AbLab AB10STMW215) and Alexa Fluor 647-conjugated anti-CD34 (BD Pharmigen 560230) were used for double positive staining of quiescent satellite cells. Satellite cells were sorted using a FACS Aria II (BD) and processed for RNA-seq and ATAC-seq. This metadata sheet includes data from the 26 ATAC-Seq samples of this project.