bioimagesUnknownMelpomeni PlataniThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-15171561JCB

Figure 4. Analysis of Cajal body antigens in CBs and mini-CBs. Fluorescence micrographs of HeLaGFP-coilin cells immunolabeled with antibodies specific for known Cajal body antigens. Maximum intensity projection images of deconvolved data sets are shown with GFP-coilin in green (A, C, E, and G) and either fibrillarin (B), Sm proteins (D), SMN (F), and SIP1 (H) in red. Fibrillarin and the Sm proteins (B and D) are detected in CBs (magenta arrowheads) but not in all of the mini-CBs (blue arrowheads). SMN and SIP1 (F and H) are detected in both CBs (magenta arrowheads) and mini-CBs (blue arrowheads). Bar, 5 µm.

Figure 7. Differential partition of components after separation of a Cajal body. (A) A plasmid containing YFP-fibrillarin was transfected into HeLaGFP-coilin cells (see Materials and Methods). Maximum intensity projections from time-lapse three-dimensional data sets are shown. Top two rows show GFP-coilin (a–c) and YFP-fibrillarin (d–f). The boxed region is enlarged in the bottom three rows to show a CB splitting that generates an unequal distribution of YFP-fibrillarin to the two resulting Cajal bodies

Figure 5. In vivo time-lapse three-dimensional imaging of HeLaGFP-coilin cells. (A) A time-lapse sequence of the GFP signal from a nucleus of an HeLaGFP-coilin cell. 12 out of a total of 60 time points are shown as a montage. Data were collected every 3 min for 2.5 h. Each image is a maximum intensity projection of 24 optical sections (0.5 µm each) spanning the entire cell nucleus. Both CBs (and mini-CBs) were observed moving. The magenta arrowhead points to a pair of Cajal bodies that fuse in the last time point. Note the changes in intensity that occur in the smaller Cajal body. The blue arrowhead points to a mini-CB that splits off from a larger CB in the second time point and then moves upwards through the nucleoplasm.

Figure 6. In vivo time-lapse three-dimensional analysis of an HeLaGFP-coilin cell nucleus showing movement of CBs through the nucleoplasm and separation of mini-CBs from Cajal bodies. 12 three-dimensional time-lapse images out of a total of 22 are shown as a montage. The image at each time point is a maximum intensity projection of 20 optical sections (0.5 µm each) scanning the entire cell nucleus. Data were collected every 3 min over a period of 2.25 h.

bioimagesFig 4 - E and FFig 6 - NoneImage 141 (Fig 6 - None)Image 137 (Fig 4 - E and F)Image 142 (Fig 7 - A)Fig 7 - AImage 136 (Fig 4 - A and B)Fig 5 - AFig 7Fig 6Fig 4 - A and BFig 5Fig 4Fig 4 - G and HFig 4 - C and DImage 135 (Fig 4 - C and D)Image 138 (Fig 4 - G and H)Image 139 (Fig 5 - A)Angus I. LamondJason R. SwedlowIlya GoldbergMelpomeni PlatanifalseIn Vivo Analysis of Cajal Body Movement, Separation, and Joining in Live Human Cells2000-12-25T11:16:57Z2018-11-29T11:16:57Z2018-11-29T10:00:54ZS-JCBD-1517156110.1083/jcb.151.7.1561