Insulin-Like Growth Factor Binding Protein-6 Alters Skeletal Muscle Differentiation of Human Mesenchymal Stem Cells

Figure 5

PMSCs treated with IGFBP-6 increased pluripotency-associated and muscle differentiation markers. IGFBP-6, OCT4, SOX2, Pax3/7, MyoD, MyoG, and MHC protein levels were quantified within PMSCs grown in muscle differentiation media with or without IGFBP-6 (375 ng/mL) supplementation using Western blots. (a) IGFBP-6 treatment increased IGFBP-6 levels as compared to PMSCs grown in muscle differentiation media only. IGFBP-6 treatment also increased pluripotency-associated markers (b) OCT4 and (c) SOX2 levels. (d) IGFBP-6 treatment increased muscle lineage commitment marker Pax3/7 at each time point. Muscle differentiation markers (e) MyoD, (f) MyoG, and (g) MHC levels increased with IGFBP-6 treatment at early time points (1–3 days) but showed reduced levels at later time points (7–14 days). Protein levels were quantified by densitometry and normalized to β-actin. Data is presented as the mean ± SEM of 3 independent experiments. Two-way ANOVA with Bonferroni’s multiple comparison test was performed to determine , , and comparing treatments, or ^#, ^##, and ^### comparing the same treatment over time.

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