ORIGINAL RESEARCH

Impact of р53 modulation on interactions between р53 family members during НаСаT keratinocytes differentiation

About authors

1 Orekhovich Institute of Biomedical Chemistry, Moscow, Russia

2 RMA “Perspektiva”, Novosibirsk, Russia

Correspondence should be addressed: Alexander L. Rusanov
Pogodinskaya 10, bld. 8, Moscow, 119121; moc.liamg@vonasur.l.rednaxela

About paper

Funding: the study involving p53 gene knockdown, ELISA and PCR tests was performed as part of the Fundamental Scientific Research Programs of the State Academies of Sciences for 2013–2020; experiments with Nutlin-3a were carried out by RMA “Perspektiva” and supported by RFBR, project № 18-44-540031/19.

Author contribution: Luzgina NG, Rusanov AL — study concept; Romashin DD, Kozhin PM, Luzgina NG, Rusanov AL — study design and literature analysis; Romashin DD, Kozhin PM, Karagyaur MN — study planning and execution; Kozhin PM, Romashin DD, Luzgina NG, Rusanov AL — data analysis and interpretation; Kozhin PM, Romashin DD — manuscript writing; Kozhin PM, Romashin DD, Karagyaur MN, Luzgina NG, Rusanov AL — manuscript editing, preparation of the final version of the article.

Compliance with ethical standards: the study was carried out in accordance with the World Medical Association Declaration of Helsinki.

Received: 2020-11-27 Accepted: 2020-12-14 Published online: 2020-12-26
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Fig. 1. Effects of TP53 modulation by knockdown and exposure to Nutlin-3a. А. Semiquantitative analysis of р53 in wild-type cells (WT) and ТР53-knockdown cells (sh). B. Metabolic activity of cells (МТТ assay) after exposure to Nutlin-3a. * significant differences between groups WT and sh (p < 0.05)
Fig. 2. Expression of differentiation markers in HaCaT cells. A. Expression of IVL, KRT10, KRT14, CASP14, TGM1. B. Immunofluorescence microscopy: KRT5 (red), KRT10 (green), DAPI (blue, nuclei), х400 magnification. C. Generalized diagram of the cells’ relative fluorescence intensities. * significant differences between treated cells (sh, Nutlin-3a) and intact cells (WT K), p < 0.05
Fig. 3. Alterations in p63 isoforms’ expression. A. Immunofluorescence microscopy: dNp63, TAp63 (red), DAPI (blue), х400 magnification. B. Generalized diagram of relative fluorescence intensities of cytoplasm and nucleus. C. Expression of TP63 isoforms (dNp63, TAp63). * significant differences between treated cells (sh, Nutlin-3a) and intact cells (WT K), p < 0.05
Table 1. Primers