Fig. 5

Aggregation of condensed chromosomes near nuclear envelope at the onset of nuclear envelope breakdown (NEB) (upper panel) and displacement of centrosomes from spindle poles (lower panel). a-c Condensed chromosomes are localized near the nuclear envelope. a, b, c –merged confocal images including the entire volume of the nucleus,a1,b1,c1 – single optical slices from central part of each nucleus. Note in cthat microtubules visualized with TPX2 encircle the nucleus and also appear in the processes. For an animated 3D reconstruction of (a) see Additional file 5A. d Disappearance of lamin A/C (lamin) immunoreactivity in the nucleus with chromosomes aggregated near the nuclear envelope. Note that some parts of chromosomes are located outside of the envelope profile (arrow), indicating NEB. Note also that nuclei of neighboring cells (probably neurons) do not show lamin A/C immunoreactivity (asterisks), whereas the nucleus of the astrocyte marked with a star displays lamin A/C immunostaining. e In arrested mitosis lamin A/C immunoreactivity has a punctate appearance distributed over the cytoplasm of the cell body. (g,h) Displacement of centrosomes (identified with gamma-tubulin [γ-tub] and pericentrin [PC], arrows) from spindle poles into the proximal parts of processes. For an animated 3D reconstruction of (g) see Additional file 5G. i-k Displacement of centrosomes (arrows) away from spindle poles. i′ shows two channels γ-tub and DAPI from three shown in i. (j) Note that centrosomes (arrows) are aside from poles. k GLAST immunostaining delineates the cell body of the astrocyte. For an animated 3D reconstruction of (i, j) see Additional files 5 I, J. l, m Several pole-like foci produced by kinetochore microtubules. Note multiple foci of gamma-tubulin immunoreactivity in (m). For an animated 3D reconstruction of (l, m) see Additional files 5 L, M. n, o NUMA immunoreactivity was predominantly located in the areas of kinetochore microtubule convergence in the spindle pole-like structures. Note that centrosomes (arrow and double headed arrow) are not associated with high immunoreactivity for NUMA and that centrosomes (double headed arrows) are displaced from spindle poles. In o′, NUMA and Nissl are shown without γ-tub. For an animated 3D reconstruction of (n) see Additional file 5 N. All images were obtained with confocal microscope. Scale bars:  a,c = 15 μm, b,d,e = 7 μm, g-n = 10 μm, o = 6 μm