PCM1 can be used to isolate the myonuclei population from whole skeletal muscle tissue.

(A) Nuclei from whole muscle lysate stained with PCM1 antibody (green) and Hoechst to visualize DNA (blue). Scale bar 20 μm. (B) Representative scatterplot showing identification of single nuclei from skeletal muscle tissue by flow cytometry. (C-D) Representative scatterplot of nuclei labeled with PCM1 and IgG, respectively. (E) Representative scatterplot of co-localization between nuclei expressing H2B-GFP and are positive for PCM1 in tibialis anterior (TA) from mouse expressing the H2B-GFP construct under the control of ACTA1, analyzed by flow cytometry. F) Co-localization between PCM1 and GFP positive nuclei in TA in the transgenic mouse model (n = 3), error bars SEM. (G) Workflow used to isolate the myo-specific nuclei from skeletal muscle. (H-J) Representative histograms of nuclear distribution and magnetic sorting efficiency for the three muscles TA, EDL and soleus (SOL) from mice by flow cytometry. (K) Quantification of nuclear distribution in full tissue (n = 3). (L) Quantification of sorting efficiency (n = 3).