Fig. 5: Activation of Peli1 expression causes epidermal hyperplasia and leads to aberrant cell cycle progression at S-G2/M phases.
a Adult rtTA control mice and rtTA-Peli1 inducible transgenic mice were treated with doxycycline (2 mg/mL) for 12 weeks. Representative immunofluorescence images of skin sections showing the expression of keratinocyte differentiation markers (keratin 14, keratin 10, and loricrin), a proliferation marker (Ki67), dermal infiltrated immune cell markers (CD3 and F4/80), and an angiogenesis marker (CD31). The dotted line indicates the border between the epidermis and dermis. Scale bars, 50 μm. b Primary keratinocytes were isolated from rtTA or rtTA-Peli1 mice after 24 weeks of doxycycline treatment. Single-cell suspensions of primary keratinocytes were fixed and stained with propidium iodide for DNA analysis via flow cytometry. Summary data are shown. Data are presented as the means ± SEMs (n = 3). *P < 0.05, ** P < 0.01, two-way ANOVA. c Double immunofluorescence staining for PCNA (red) and phospho-H3S10 (green) expression in skin sections of rtTA and rtTA-Peli1 mice after 24 weeks of doxycycline treatment. Scale bars, 50 μm (inset magnification, ×630). d Quantification of positively stained keratinocytes in skin sections. PCNA: a marker of S phase; phospho-H3S10: a marker of mitosis; PCNA and phospho-H3S10 double-positive: a marker of G2 phase. The results are presented as the means ± SEMs (n = 5). e Immunoblot analysis of the indicated protein in skin tissues derived from rtTA and rtTA-Peli1 mice after 24 weeks of doxycycline treatment.
