Figure 3.

Fisetin treatment of cell co‐culture system of Z24^−/− FAPs and WT MPCs. (A) BrdU proliferation assay and myogenesis assay showed that the proliferation and myogenesis potentials of WT MPCs were decreased when being co‐cultured with Z24^−/− FAPs; while the treatment of the co‐culture system of WT MPCs and Z24^−/− with fisetin effectively rescued the proliferation potential of the WT MPCs; also, fisetin treatment of WT MPCs or co‐culture system of WT MPCs and WT FAPs did not affect the proliferation and myogenesis potentials of WT MPCs. Immunofluorescent staining of f‐MHC (fast‐type myosin heavy chain) in WT MPCs was performed to compare myogenesis potential (myotube formation) in the co‐culture system. (B) Schematic presentation of the transwell co‐culture system with WT FAPs or Z24^−/− FAPs being cultured in the upper chamber and WT MPCs being cultured in the lower chamber. (C) Quantitation of proliferation and myogenesis of the different groups shown in (A). The quantification of myotube formation was presented by the fusion index of muscle cells, which is the number of nucleus incorporated into multinucleate myotubes (f‐MHC+), compared with total number of nuclei. Data analysis was performed with random pictures from each group of cells, and the sample number here N = 6 (three biological replicates with two technical replicates for each).