Il 36α

Using formalin-fixed, paraffin-embedded specimens, hematoxylin and eosin and immunohistochemical staining were performed using antibodies against IL-36α(R&D Systems, Minneapolis, MN, USA), IL-36Ra (R&D Systems), nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-κB; Abcam, Cambridge, UK), IL-23 (Biolegend, San Diego, CA, USA), IL-17 (Abcam), and IL-8 (Abcam). The immunohistochemical results were analyzed individually in the epidermis and dermis using Image Pro Plus Version 4.5 (Media Cybernetics Co., Silver Spring, MD, USA). The expressions of each immunohistochemical staining were determined by ratio of the stained area to measured epidermal area and dermal area on the representative area of each specimen as a blinded state. Dermal area was defined as the area within 200 µm from epidermo-dermal junction and representative area was defined as the area represents average stained area in consideration of overall tendency at each specimen. To determine the expression of NF-κB positivity, the ratio of NF-κB-positive cell count to the overall cell count within the epidermis or dermis was determined. For the NF-κB staining, positive findings were defined as strong nuclear staining, easily observed at the scanning magnification. For each frame, the tracing was repeated 3 times and the mean was determined.