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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 4. — Plk4 over-expression leads to hyperproliferation and abnormal differentiation of cells in suprabasal layers. (a) Immunofluorescence of cryosections of P2 back skin from mice of indicated genotypes treated with doxycycline as indicated (+DOX). Note: Ki67 reveals proliferating cells in basal and suprabasal (white asterisk) epidermal layers of Plk4^OE/Plk4^OE; p53^KO/p53^KO (+DOX) mice; K6, restricted to hair follicles in controls, is present in epidermis following Plk4 induction (arrowheads). Basal layer marker K5 present throughout epidermis following Plk4 induction; distribution of early differentiation marker, K10 not affected and does not reveal differences between the experimental samples and the control at this stage. B, basal; Der, dermis; Epi, epidermis; Sp, suprabasal; dotted lines, dermo-epidermal border. (b) Quantitative RT-PCR assays for levels of indicated transcripts in total back skin from 2 day pups of indicated genotypes and treatment (n = 3 and three replicates for each sample). Note: elevated Plk4 transcripts consistently correlate with low expression of late differentiation genes filaggrin (Fla), involucrin (Inv) and loricrin (Loc). (c) Quantitative RT-PCR assays as in (b) for P21 (mp21) and ΔNP63. (d) Immunofluorescence analysis of back skin from 20 day pups of indicated genotypes and treatment. Note: Ki67-positive cells in basal and suprabasal cells (arrowheads) and K6 in multiple layers co-localizing with K5 (white arrow) in Plk4^OE/Plk4^OE; p53^KO/p53^KO (+DOX) mice; expanded staining of K5 and K10. (e) Ratio of basal to suprabasal cells indicated as mean ± s.d. (f) Proportion of cycling cells in epidermis. Mean ± s.d. % of cells showing Ki67-positive immunostaining. Significance determined by Student's t-test. *p < 0.05.