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The medium was replaced, and after 8 h the cell supernatant was collected and co-incubated with RAW264.7 cells for 4 h

After washing with PBST (0.05% Triton X-100), secondary antibodies (anti-mouse Alexa Fluor 488 (1:500) and anti-rabbit Alexa Fluor 594 (1:500)) were stained at 37 C for 1 h, and nuclei were stained with DAPI for 5 min

On the other hand, Langerhans cells are increased in the epidermis, the basement membrane is thickened, and cytoplasmic vacuolization phenomena are observed.44,45 It should be noted that Dickkopf 1 (DKK1) protein, involved in reducing melanogenesis, is overexpressed in fibroblasts in lesional skin.46 There is also increased fibronectin expression and reduced elastin in lesional dermis, with no changes observed in collagen fibers.47,48 Genetic changes involved in the etiopathogenesis of vitiligo Vitiligo is a complex disease with a significant genetic component, estimated between 75% and 83%

43 , e47e47 (2015)