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Actin labeling reveals the differentiation state of the myotubes. C2C12 cells are plated on IPN patterns, grown for 2, 6, and 14 days, and then stained for F-actin (red), and DNA (blue). Arrows in the day 2 images highlight clear membrane divisions between cells, showing that the myoblasts have not yet fused. Although the cells are clearly fused by day 6, actin maintains a stress fiber-like orientation that persists thereafter.
