GSM8254122: WT4_MuSCs-ATAC; Mus musculus; ATAC-seq
Muscle Stem Cells (ITGA7+/VCAM1+/CD11b-/CD45-/CD31-/Ly6a-) were isolated by Fluorescence Activated Cell Sorting (FACS) into ATAC-Seq lysis buffer (10 mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 0.1% NP-40, 0.01% Digitonin). EDL muscle was dissected and myofibers were isolated as previously described (Blackburn, D.M. et al. Journal of Biological Chemistry, 2019). After isolation and selection of a single myofiber, myofibers were permeabilized with 0.5% Triton X-100 in PBS  for 15 minutes at room temperature (RT). The permeabilization step was followed by two washes with PBS1X. The ATAC-Seq was then performed as previously described (Sahinyan K. et al. eLife, 2022 , Sahinyan K. et al. bioProtocol, 2022). ATAC-Seq libraries were prepared as previously described (Sahinyan K. et al. eLife, 2022 , Sahinyan K. et al. bioProtocol, 2022) with incorporation of Illumina Nextera XT adaptors.