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. 2021 Mar 31;22(7):3645. doi: 10.3390/ijms22073645

Figure 2.

Figure 2

Morphological analyses of formalin-fixed and paraffin-embedded tissue sections from control EDL muscles, EDL muscles damaged by ex vivo forced eccentric contraction (EC) and EDL muscles damaged by EC in the presence of bone marrow-mesenchymal stromal cell (MSC) conditioned medium (EC + MSC-CM). (a–c) Representative confocal immunofluorescence images of muscle tissue sections immunostained with antibodies against cytochrome (Cyt) c (red) and counterstained with the membrane dye wheat germ agglutinin (WGA) (green). (d) Quantitative analysis of the percentage of myofibers with Cyt c fluorescence intensity (FI) higher than that of control. (e–g) Representative superimposed confocal immunofluorescence and differential interference contrast (DIC, grey) images of muscle tissue sections immunostained with antibodies against the pro-apoptotic marker activated/cleaved caspase-3 (green). (h) Quantitative analysis of the percentage of cleaved caspase-3+ myofibers. (i–k) Representative confocal immunofluorescence images of muscle tissue sections immunostained with antibodies against the pro-survival/anti-apoptotic factor AKT (green) and counterstained with the nuclear dye propidium iodide (PI, red). Insets: representative confocal immunofluorescence images of muscle tissue sections immunostained with antibodies against the activated phosphorylated form of AKT (p-AKT, green) and counterstained with PI (red). Images are representative of at least 3 sections from each of 4 control, 4 EC and 4 EC + MSC-CM muscles. Scale bars = 25 μm. (l) Quantitative analysis of AKT FI. a.u.: arbitrary units. Values are mean ± SEM. ** p < 0.01 vs. control; # p < 0.05 and ## p < 0.01 vs. EC.