Figure 3

Chrysotile treated cells during cell cycle. Laser scanning confocal microscope images of HK2 cells treated with chrysotile for 48 h and recovered for 24 h in fiber-free culture medium: A) metaphase cell with multipolar spindle (microtubules in blue, DNA in red, actin filaments in green), presenting small fragments of chrysotile (pink) inside; A') and A") orthogonal projections showing the intracellular localization of chrysotyle fibers; B) 3D reconstructions of optical sections of the same cell; small chrysotile fibers are also seen outside the cell (small pink dots). Chrysotyle treated HK2 cell in late telophase with a chrysotyle fiber between the two daughter cells: C) Image of nuclei in red and crysotyle fibers in pink, D) merge with microtubules (green) image; the microtubules seem to be normally organized, forming the midbody in cytokinesis region; E) the 3D reconstruction of their optical sections evidence the cell- fiber interaction, with the fiber inside the cell and between the daughter nuclei (red). G and F) Confocal image showing four centrosomes evidenced by anti-gamma tubulin (green) and condensed mitotic chromosomes (red). Chrysotile fibers (pink) are observed inside the cell in confocal image. The 3D reconstruction (G) evidence the fiber involved by chromatin.