After 4 h, the medium containing Lipofectamine was replaced with a fresh regular medium. a longer time scale, example 2. DKO neural stem cells (NSCs) were induced to differentiate into neurons. At 3 days postdifferentiation, the cells were stained with SiR-Hoechst and subjected to live imaging by confocal microscopy. Images were taken at 30-min intervals for 44 h. Z-series of 25 focal planes with a step size of 0.5 m were acquired. Cells actively migrated during the imaging period. Thus, the nucleus was centered to make it easier to follow. The maximal projected image sequence shows dynamic nuclear deformation and chromocenter (CC) clustering. Scale bars: 10 m.(MOV) pcbi.1007289.s003.mov (252K) GUID:?17781591-B25E-40FC-BA22-B103840AE8F5 S3 Movie: Live imaging of double-knockout (DKO) neuronal cells at 3 days postdifferentiation on a longer time scale, example 3. DKO neural stem cells (NSCs) were induced to differentiate into neurons. Col003 At 3 days postdifferentiation, the cells were stained with SiR-Hoechst and subjected to live imaging by confocal microscopy. Images were taken at 30-min Col003 intervals for 44 h. A Z-series of 25 focal planes with a step size of 0.5 m were acquired. Cells actively migrated during the imaging period. Thus, the nucleus was centered to make it easier to follow. The maximum projected image sequence shows dynamic nuclear deformation and chromocenter (CC) clustering. Scale bars: 10 m.(MOV) pcbi.1007289.s004.mov (192K) GUID:?7833E1D3-871A-4D3E-92F7-57F87AD1F399 S4 Movie: Results of numerical simulation of chromocenter (CC) clustering by dynamic nuclear deformation. The period of simulation is approximately 33 h.(MOV) pcbi.1007289.s005.mov (2.1M) GUID:?A9F4681E-9EA4-4513-9702-947B43AF4952 S5 Movie: Live imaging of actin dynamics in double-knockout (DKO) neuronal cells at 3 days postdifferentiation, example 1. DKO neural stem cells (NSCs) were induced to differentiate into neurons. At 3 days postdifferentiation, the cells were stained with SiR-Actin and Vybrant DyeCycle Orange Stain and Col003 subjected to live imaging by confocal microscopy. Images were taken at 15-min intervals for 3 h. A Z-series of 77 focal planes with a step size of 0.2 m were acquired. The maximum projected image sequence shows dynamic actin movement. In this movie, three images are connected horizontally. Left, middle, and right side of images represent merged, nucleus (Vybrant DyeCycle Orange Stain), and actins (SiR-Actin), respectively. Scale bars: 10 m.(MOV) pcbi.1007289.s006.mov (2.0M) GUID:?C87FE32A-DBB8-4274-8C00-AE54EB60B655 S6 Movie: Live imaging of actin dynamics in double-knockout (DKO) neuronal cells at 3 days postdifferentiation, example 2. DKO neural stem cells (NSCs) were Rabbit Polyclonal to CHST10 induced to differentiate into neurons. At 3 days postdifferentiation, the cells were stained with SiR-Actin and Vybrant DyeCycle Orange Stain and subjected to live imaging by confocal microscopy. Images were taken at 15-min intervals for 3 h. A Z-series of 77 focal planes with a step size of 0.2 m were acquired. The maximum projected image sequence shows dynamic actin movement. In this movie, three images are connected horizontally. Left, middle, and right side of images represent merged, nucleus (Vybrant DyeCycle Orange Stain), Col003 and actins (SiR-Actin), respectively. Scale bars: 10 m.(MOV) pcbi.1007289.s007.mov (2.6M) GUID:?2E147B3B-1748-49CB-B8E6-81A2B1202518 S7 Movie: live imaging of P15 mouse rod cells. Retinal tissue was excised from P15 mouse, stained with Hoechst 33342 and subjected to live imaging using two-photon microscopy. Images were taken at 10-min intervals for 3 h. Z-series of 103 focal planes with a step size of 0.2 m were acquired. Center of mass in chromocenter (CC) clusters of some cells are represented by colored balls. Color coded lines represent trajectories of CC clusters. Scale bar: 5 m.(MOV) pcbi.1007289.s008.mov (9.1M) GUID:?CC9942A7-3656-462C-8746-A9D965305F14 S8 Movie: Dynamics deformation with affinity between heterochromatin and nuclear envelop. (MOV) pcbi.1007289.s009.mov (645K) GUID:?98814288-8CC0-4C53-AD6B-FFC3EE319313 S1 Fig: Establishment of DKO cell lines. (A) The strategy for targeted gene inactivation for establishment of the double knockout (DKO) cell lines is shown in Col003 the left panel. Grey and black boxes indicate untranslated regions and coding sequences, respectively. The right-hand.