JCS016907 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Latrunculin A-treated hepatocytes elongate with suppression of cortical rigidity. Hepatocytes were isolated from 25-day-old rat (after weaning) and cultured in serum-free medium with epidermal growth factor (5 nM), insulin (100 nM) and dexamethasone (25 nM). Hepatocytes were mounted on time-lapse videomicroscopy 60 hours after plating. Mitosis of mononucleated 2n cells were monitored in the presence or absence of latrunculin A (added 1 hour before imaging). Cell elongation from metaphase to telophase (mean increase in cell length from metaphase ± standard error) was determined using time-lapse sequences. Data are presented as the mean±s.e. from ten independent time-lapse sequences.

• Supplemental Figure S2 -

  Fig. S2. Quantification of phospho-Myosin light chain II along the cortex during completed or incompleted cytokinesis processes. Phospho-Myosin light chain II immunofluorescence was quantified along the cortex in early telophase of complete (left panel) and incomplete (right panel) cytokinesis. Quantification was measured only on half of the cell, from one pole to the other (see Fig. 4B). Fifty early telophases were analyzed in four independent cultures.

• Supplemental Figure S3 -

  Fig. S3. Quantification of microtubule network at the equatorial region during completed or incompleted cytokinesis processes. Hepatocytes were isolated from 25-day-old rat (25±3.6% of mitotic hepatocytes incomplete cytokinesis). Tubulin immunofluorescence was quantified at the equatorial region of hepatocytes that complete or incomplete cytokinesis (see Materials and Methods). Fifty anaphases/telophases were analyzed in four independent primary hepatocyte cultures. Bars represent s.e., P<0.0001. Microtubule density significantly increased, between anaphase and telophase, only in hepatocytes that completed cytokinesis.

• Supplemental Figure S4 -

  Fig. S4. Aurora B, Prc1 and Plk1 behaviour during incomplete cytokinesis process. Hepatocytes were isolated from 25-day-old rat. Sixty hours after plating, hepatocytes were fixed and stained with anti-Aurora B (red) or anti-Prc1 (red) or anti-Plk1, and with anti-β-tubulin (green) antibodies. Nuclei were stained with Hoechst 33342. As described for MgcRacGAP, Aurora B, PRC1 and Plk1 only localized to the remaining interdigitating microtubules during incomplete cytokinesis. Bars, 5 μm.

• Movie 1 -

  Movie 1. Before weaning, division of diploid hepatocyte gives rise solely to two diploids progenies.

• Movie 2 -

  Movie 2. After weaning, division of diploid hepatocyte gives rise to a tetraploid progeny (binucleated 2×2n).

• Movie 3 -

  Movie 3. Before weaning, diploid hepatocyte treated with HA1077 present an anaphase cell elongation defect leading to the genesis of binucleated 2×2n hepatocyte.