Journal: Scientific Reports

Article Title: Foxn1 expression in keratinocytes is stimulated by hypoxia: further evidence of its role in skin wound healing

doi: 10.1038/s41598-018-23794-5

Figure Lengend Snippet: Foxn1 regulates keratinocyte differentiation and migration. Flow cytometric analysis of keratinocytes: non-transfected and transfected with Ad-Foxn1 or Ad-GFP, showing the percentage expressing keratin 14 (K14) only, keratin 10/keratin 14 (K10/K14) and K10 only ( a ). Representative Western blots of keratin 16 and involucrin protein expression in keratinocytes transfected with Ad-Foxn1 or Ad-GFP (control) ( b ). Full-length blots are presented in Supplementary Figure . Migratory abilities after pre-treatment with mitomycin C and wounding in monolayer cultures were analysed in non-transfected B6 keratinocytes and in keratinocytes transfected with Ad-Foxn1 or Ad-GFP ( c , d ). Representative images were taken at 0, 20, 32 and 68 h ( d ). Dotted lines indicate the distance between migrating keratinocytes ( d ). Migration is expressed as the percentage of the distance between the unclosed edges ( c ). Each well was prepared in duplicate, and the experiment was repeated two times (n = 11 animals). Values are the mean ± SD. Asterisks indicate significant differences (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).

Article Snippet: Keratinocytes were trypsinized 48 hours after transfection, suspended in warm PBS and incubated for 30 min with the following antibodies: anti-keratin 10-PerCP (mouse monoclonal, clone SPM261; Novus Biologicals conjugated with PerCP; Lynx Rapid PerCP antibody conjugation kit; AbD Serotec) and anti-keratin 14-APC (mouse monoclonal, clone LL002; Novus Biologicals conjugated with APC; Lynx Rapid APC antibody conjugation kit; AbD Serotec).

Techniques: Migration, Transfection, Expressing, Western Blot, Control