1. Skin-Specific Deletion of Mis18α Impedes Proliferation and Stratification of Epidermal Keratinocytes 
Koog Chan Park, Minkyoung Lee, Yoon Jeon, Raok Jeon, Sung Hee Baek, Ho Lee, Keun Il Kim 
Journal of Investigative Dermatology 
Volume 137, Issue 2, Pages (February 2017)
DOI: /j.jid
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2. Figure 1
Skin-specific disruption of Mis18α in mice. (a) In situ hybridization of Mis18α mRNA. Anatomical view (left), autoradiography detection (bright labeling in middle), and control hybridization (right) are shown. Scale bars at E9.5 = 1 mm, at E12.5 = 1 mm, at E15.5 = 0.5 cm, at P1 = 1 cm and at P10 = 1 cm. (b) The strategy for the skin-specific deletion of Mis18α and an example of the genotyping results. A (–) mark indicates whole-body deletion of Mis18α. P1, P2, and P3 are primers for genotyping. (c) Summary of genotypes of offspring from Mis18α+/−;K14-Cre and Mis18αf/+;K14-Cre breeding. Numbers in parentheses indicate percentage survival. (d) Representative pictures of newborn mice. (e) Acidic X-gal dye-penetration assay with newborn mice (left). Blue color indicates the accumulation of X-gal inside the skin. Western blot analysis of epidermal keratinocytes from newborn mice for the proteins involved in tight-junction formation and in the terminal differentiation (right). E, embryonic day; K, keratin; P, postnatal day.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions

3. Figure 2
Defective stratification of epidermis in Mis18α-deficient mice. Sections of the dorsal and ventral skin from newborn wild-type and Mis18αf/−;K14-Cre mice were subjected to either hematoxylin and eosin staining (upper panel) or immunostaining with antibodies against keratin-14, keratin-10, loricrin, and filaggrin (lower panels). DAPI-stained nuclei are shown in blue. Scale bar = 200 μm. H&E, hematoxylin and eosin; Krt, keratin.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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4. Figure 3
Analysis of cell proliferation and apoptosis in the epidermis. Sections of the dorsal skin from newborn mice were (a) stained with anti-Ki67 antibody and (b) subjected to TUNEL staining. Red signal indicates TUNEL-positive cells, and blue signals represent DAPI-stained nuclei. White line represents the basal layer. Scale bar = 200 μm. (c) Histogram represents the percentage of TUNEL-positive cells per epidermis of a newborn mouse. The number of TUNEL- and DAPI-positive cells in any three distinct fields of equal area was counted for every section. Three mice were used in each group. Data are shown as mean ± standard deviation. ∗P < (d) Keratinocyte extracts prepared from the epidermis of newborn mice were subjected to immunoblot against indicated proteins. Red asterisk indicates nonspecific band.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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5. Figure 4
Comparison of dorsal and ventral epidermis in Mis18α-deficient skin at E15.5 and E17.5. Sections of the dorsal and ventral skin from wild-type and Mis18αf/−;K14-Cre mice (a) at E15.5 and (b) at E17.5 were subjected to H&E staining, immunostaining with antibodies against keratin-14 and keratin-10, and TUNEL staining. DAPI-stained nuclei are shown in blue in a and b. Scale bar = 200 μm. (c) K14-Cre–mediated activation of the Rosa26 β-galactosidase reporter (Rosa26R) was monitored at E11.5. K14-Cre mice were crossed with the Rosa26R mice, and embryos at E11.5 were stained for β-galactosidase activity (X-gal). Scale bar = 100 μm. E, embryonic day; H&E, hematoxylin and eosin; Krt, keratin.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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6. Figure 5
Comparison of Cenp-A protein levels in keratinocytes. (a) Immunocytochemistry images were obtained from in vitro culture of keratinocytes prepared from the skin of newborn, wild-type, or Mis18αf/−;K14-Cre mice. Scale bar = 20 μm. (b) The amount of Cenp-A protein in keratinocytes prepared from the skin of newborn, wild-type, or Mis18αf/−;K14-Cre mice was compared by immunoblotting. α-Tubulin was used as loading control.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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7. Figure 6
The effects of Mis18α depletion on chromosome stability and DNA damage responses in HaCaT cells. HaCaT cells were infected with the lentivirus expressing control short hairpin RNA (shControl) or short hairpin RNA against Mis18α (shMis18α). (a) Chromosomes (DAPI-stained as blue), microtubules (stained with anti-α-tubulin antibody, red), and CENP-A (green) were visualized after 7 days of infection. Open arrows indicated mitotic cells. (b) Mitotic cells in a were counted for chromosome misalignment (control cells, n = 38 vs. Mis18α knockdown cells, n = 41). Blue bar represents cells showing correctly aligned mitotic chromosomes, and red bar represents cells having misaligned chromosomes.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions