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Figure 5: Characteristics of EMT in shikonin-treated wound healing in mouse skin tissues. (a) Wound closure rates of skins subjected to different treatments (shikonin versus control). The data represent the mean ± SEM of six mice. At 2 d after treatment, histological and immunohistochemical characteristics of acetone- (1) and shikonin-treated skins (2) were analyzed. Long arrows indicate the original edges of each wound. The dotted line in panel 2 indicates the terminally damaged boundary line between the epidermal and dermal layers. (b) Tissue biopsy samples from each test group were also stained for the expression of E-cadherin (red), FSP1 (green), and DAPI (blue). Triangle arrows indicate the presence of mesenchymal cells in the epidermis (3 and 4). The presence of vimentin (red) and FSP1 (green) double-stained cells (yellow and indicated with arrowheads) further indicates that these epithelial cells have undergone the EMT process in test skin tissues (5 and 6). (c) Changes in expression levels of mesenchymal-associated microRNAs, including mir-200a, mir-200b, mir-200c, mir-141, mir-429, and mir-205, as well as the Zeb1, Zeb2, and E-cadherin mRNAs (d) in shikonin-treated skin wound, as measured by real-time PCR analyses. The significance of differences was analyzed by one way ANOVA ( and ).