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. Author manuscript; available in PMC: 2018 Mar 11.
Published in final edited form as: Nat Cell Biol. 2017 Sep 11;19(10):1189–1201. doi: 10.1038/ncb3605

Figure 1. Visualizing nuclear movement to the periphery.

Figure 1

A. Timeline of muscle differentiation in the in vitro system used to study peripheral nuclear positioning and transversal triad formation. Nuclei are in red, myofibrils in white (with z-lines in green) and transversal triads as purple lines. Day 3: myofibril formation. Day 5: initiation of peripheral nuclear positioning. Day 7: transversal triad formation.

B. Kymograph from a time-lapse movie of a 5-day myofiber depicting peripheral movement of a nucleus (H2B-iRFP, red) through myofibrils (YFP-α-actinin, green). Left: view from the side, surface three-dimensional (3D) rendering. Middle: view from the right side, with transparent myofibrils 3D rendering. Right: view from the top, surface three-dimensional rendering. Time, hh:mm. Scale bar, 10 μm.

C. Two dimensional (2D) view of the central plane of a kymograph from a time-lapse movie of a 5-day myofiber depicting peripheral movement of a nucleus (H2B-iRFP, red) through myofibrils (YFP-α-actinin, green). Scale bar, 10 μm.

D. Representative image of a nucleus squeezing to the periphery from an in vivo isolated myofiber of a newborn mouse and stained for myofibrils (α-actinin, green) and nucleus (red). Left: 3D rendering. Middle left: 2D orthogonal view, yellow lines represent slices seen in right side panels. Middle right: 2D plane from yellow slice 1. Right: 2D plane from yellow slice 2. Scale bar, 10 μm. Image shown is representative of 2 experiments.

E. Representative image of a nucleus squeezing to the periphery after performing a clearing protocol of a whole muscle in a newborn mouse and stained for myofibrils (α-actinin, green) and nucleus (red). Left: 3D rendering. Middle left: 2D orthogonal view, yellow lines represent slices seen in right side panels. Middle right: 2D plane from yellow slice 1. Right: 2D plane from yellow slice 2. Scale bar, 10 μm. Image shown is representative of 2 experiments.

F. Transversal electron micrograph of a 4.5 day myofiber showing a nucleus beginning to protrude towards the periphery. Scale bar, 2 µm. “M” annotate myofibrils.

G. Transversal electron micrograph of a 4.5 day myofiber showing a nucleus during budding to the periphery. Scale bar, 500 nm. 4x Magnifications corresponding to the yellow square is shown below the image. “M” annotate myofibrils. Image shown is representative of 4 experiments.