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. 2020 Sep 3;15(4):926–940. doi: 10.1016/j.stemcr.2020.08.002

Figure 2.

Figure 2

Damaged-Myofiber-Derived Factors Activate Satellite Cells

(A) A schematic illustrating the experimental procedure. Individual myofibers were freshly isolated from EDL muscles and mechanically damaged with a Pasteur pipette. Damaged myofibers shrank immediately (arrowheads). A growth-factor-rich medium (plating medium: PM) condition was used for a positive control. Intact (undamaged) myofibers were co-cultured with damaged myofibers (co-cultured, intact myofibers:damaged myofibers = 1:1).

(B–E) Individual myofibers associated with satellite cells were cultured in DMEM with or without damaged myofibers for (B) 48 h or (D) 72 h and then immunostained for PAX7 and MYOD (quantified in C or E, respectively). (n = 4 mice per condition; >15 individual myofibers per mouse were counted.) ∗p < 0.05. Scale bars, 50 μm.

(F–I) Individual myofibers associated with satellite cells were cultured in DMEM with or without damaged myofibers for 72 h and then immunostained for PAX7 and Ki67 or PAX7 and EdU (quantified in G or I, respectively). (n = 4 mice per condition; >15 individual myofibers per mouse were counted.) ∗p < 0.05. Scale bars, 50 μm.