3D visualization of epidermal TJs and LCs. (A) TJ network visualized by anti–ZO-1 antibody in epidermal sheet demonstrated en face by confocal microscopy. Bar, 50 µm. (B) A 90° rotation image of the boxed area in A. The top shows an en face image of ZO-1 staining followed by rotation images stained for the indicated TJ molecules. Among the three layers of granular layer, SG1 to SG3, TJs (arrowheads) were found exclusively in the SG2 layer (asterisk and dashed lines), and rotation images display TJs as bright dots with intense ZO-1 and claudin-1 signals (see Video 1). (C) TJs and LCs in unperturbed skin. Z-slice numbers are indicated in each panel. ZO-1 and claudin-1 colocalized at KC–KC TJs, and LCs (MHC II⁺) coexpressed claudin-1 at high levels. (D) A 90° rotation image of the boxed area in C (see Video 2). Tips of LC dendrites (arrows) localized within TJ barriers (arrowheads). Bars, 20 µm. Data presented is representative of five mice.