Proliferation of human LCs in the epidermis from healthy controls and AD patients. Epidermal sheets were obtained from four healthy donors and two AD patients and analyzed by confocal microscopy for expression of langerin (green) and Ki67 (red). (A) Representative maximal intensity projections of Z-stacks of healthy epidermis. Ki67⁺/langerin⁺ LCs are circled in white, and nuclear localization of Ki67 was confirmed by three-dimensional visualization (for more information see Video 5). (B) Ki67⁺ LC doublets were detected in human epidermis. (C) A significantly higher percentage of Ki67⁺ LCs was observed in AD epidermis compared with healthy controls (14.5 ± 1.55% and 3.21 ± 0.33%, respectively; P = 0.003 ; n = 2 and 4, respectively). A minimum of 500 LCs, as visualized by langerin expression, were analyzed per healthy donor and patient. Results are expressed as mean percentages of Ki67⁺LCs ± SD. (D) Representative maximal intensity projections of Z-stacks of epidermis from AD patients. A group of three Ki67⁺ LCs is shown as a three-dimensional reconstruction. Ki67⁺/langerin⁺ LCs are circled in white. The scale is graduated in micrometers.