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. 2015 Dec 23;5(12):150209. doi: 10.1098/rsob.150209

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© 2015 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 1. — Tumour formation following tetracycline-inducible conditional Plk4 expression. (a) Doxycycline associates with the rtTA that binds the TRE, leading to transcriptional activation of Plk4. (b) Tumour incidence in Plk4 transgenic mice in wild-type background (Plk4^OE/Plk4^OE) or p53 null background (Plk4^OE/Plk4^OE; p53^KO/p53^KO) with or without treatment with doxycycline (+DOX) to promote Plk4 over-expression. The differences observed between Plk4^OE/Plk4^OE; p53^KO/p53^KO (n = 24) and Plk4^OE/Plk4^OE; p53^KO/p53^KO +DOX (n = 14) survival curves are significant (**p < 0.01; Student's t-test). (c) Proportions of sarcomas and lymphomas. Note that animals that developed sarcomas also showed lymphomas. (d) Architecture of thymus and lymph nodes is obliterated and replaced by sheets of large, round cells with vesicular nuclei (Vn, black arrows). Apoptotic cells were also present (Ap, black asterisk). (e) Multicentric high-grade large cell lymphoma. (f) Kidney has normal architecture but contains multifocal cortical interstitial and sub-capsular infiltrates of large lymphocytes (Li). (g) Large sheets of lymphocytes (Li) attached to pericardial surface of heart wall. (h–j) These tumours are sarcomas isolated from Plk4^OE/Plk4^OE; p53^KO/p53^KO mice. Typically, large masses of cells extended into muscle fibre bundles (M) and into attached adipose surrounding nerves and blood vessels. (i,j) Higher magnifications of sarcomas showing pleomorphic and anaplastic cells. Examples of giant (G) or multinucleated (MM) cells are indicated. These tumours were found close to the front limbs with high percentage of mitotic cells (an average of 5–10/40× field). (k) Paraffin section of samples from sarcomas where stained to reveal γ-tubulin or acetylated-tubulin (green). DNA is shown in red. Three different samples were analysed and showed a high mitotic index (8.55 ± 2.53%, n = 1400 cells/sample) in agreement with histological analysis made after H&E staining. (l) Proportion of cells that show one pair or two pairs of centrioles per cell or show centriole/centrosome amplification (more than 2 pairs of centrioles). (m) Mitotic progression in sarcomas from cryostat sections. (n) Proportions of mitotic abnormalities in sarcomas. Quantification in (l–n) performed in three different sarcomas; 500–1000 cells analysed per sarcoma.