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

Figure 5.

Figure 5

Extracellular Treatment with Metabolic Enzymes Induces the Entry of Satellite Cells into the G1 Phase

(A and B) To test whether metabolic enzymes identified in Figure 4 as DMDFs activate satellite cells, individual myofibers were freshly isolated from EDL muscles and treated with recombinant metabolic enzyme proteins in DMEM for 72 h. Immunostaining of satellite cells associated with individual myofibers for PAX7 and MYOD was performed (A, high-glucose condition; B, non-glucose condition). (Left, absolute numbers of positive cells; right, relative ratio of positive cells.) PBS, BSA, and urease were used as a negative control, a non-specific protein control, and non-muscle enzymatic control, respectively. Damaged myofibers (Damaged) were used as a positive control. All scale bars, 50 μm. Values are means ± SE (n = 3–4 mice per condition). Asterisk (∗) and pound sign (#) indicate differences compared with urease control (p < 0.05).

(C) To examine whether the HGF signaling pathway is involved in the DMDF-induced satellite cell activation, satellite cells associated with individual myofibers were treated with recombinant metabolic enzyme proteins in the presence of PHA-665752 (c-MET inhibitor) in DMEM for 72 h. Immunostaining for PAX7 and MYOD was performed (left, absolute numbers of positive cells; right, relative ratio of positive cells). Values are means ± SE (n = 4 mice per condition). Asterisk (∗) and pound sign (#) indicate differences compared with urease control (p < 0.05).