JCS00709 Movies

Files in this Data Supplement:

• Movie 1 - A movie illustrating the contractile activity of GFP-myosin expressed in embryonic chick cardiomyocyte. This cell is expressing the GFP-myosin R403Q mutant. The cell was selected in fluorescence imaging mode then recorded using high resolution DIC optics with video enhanced contrast at 30 fps. The size of the original movie has been reduced 2-fold out of bandwidth considerations. The speed of contraction and relaxation was measured by tracking the relative position of two Z-lines as illustrated in Fig. 3B. Imaging Conditions: 100´ 1.3 NA oil DIC objective recorded a Hamamatusu C2400 CCD camera and Argus-20 Image processor.
• Movie 2 - A short movie loop illustrating the spontaneous contraction of embryonic chick cardiomyocytes expressing WT GFP-myosin. This video was taken using an intensified CCD camera at 15 fps and illustrates the spontaneous contractions triggered by Ca²⁺ sparks. The cell was observed continuously beating for 9 minutes. Imaging conditions: 60´ 1.4 NA oil objective with 3.3´ intermediate lens recorded with a image intensifier coupled to a vidicon camera.
• Movie 3 - Small area of a multinucleated C2C12 myotube expressing WT GFP-myosin and illustrating the asynchronous oscillation of the sarcomeres in these myofibrils. Unlike the embryonic cardiomyocytes, coordinated contractions of the multinucleated myotubes rarely occur prior to 7 days after induction of differentiation. However, the individual sarcomeres exhibit uncoupled oscillatory motions that may be necessary for lateral alignment of adjacent myofibrils. Imaging conditions: 40´ 0.7 NA objective recorded with a cooled CCD camera at 100 ms/frame (5-6 fps).