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. 2018 Dec 13;9:5293. doi: 10.1038/s41467-018-07688-8

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Long-term TPA treatment increases BMDCs in the epidermis of the skin. a BMDCs were detected in the epidermis of the skin with different treatment groups (before TPA treatment, no treatment, acetone-treated, and TPA-treated) and specific areas in white and magenta boxes are magnified. Arrows indicate keratin-negative BMDCs and arrowheads indicate keratin-positive BMDCs. b The number of GFP-positive cells in the epidermis are compared among three different treatment groups (no treatment, acetone-, and TPA-treated) at three different time points. The treatment types (no treatment, acetone, and TPA) are different across all weeks (P = 5.35 × 10E−11). The average time effects are different across all treatment levels (P = 1.32 × 10E−11). Additionally, the treatment type and time interact in a non-additive fashion (P = 6.6 × 10E−10). (n = 3, over all P value = 5.72 × 10E−13 as determined by the F-statistic, mean ± s.d.). c Keratin-positive BMDCs were observed in the basal layer, the suprabasal layer, and hair follicle. Areas in white boxes are magnified or shown in single color. Arrows indicate keratin negative BMDCs and arrowheads indicate keratin-positive BMDCs. d The number of GFP/Keratin-positive (double positive) BMDCs in the epidermis are compared among three different treatment groups (no treatment, acetone-, and TPA-treated) at three different time points. The number of double positive cells are increased in long-term TPA-treated groups. Both treatment type (acetone and TPA) and time (in weeks) have significant effects. This implies that the means of treatment types (no treatment, acetone, and TPA) have different effects across all weeks (P = 3.87 × 10E−08). The average time effects are different across all treatment types (P = 1.95 × 10E−07). Finally, there is a significant interaction term; this means that treatment type and time interact in a non-additive fashion (P = 1.53 × 10E−07). (n = 3, overall P value = 1.22 × 10E−09 as determined by the F-statistic, mean ± s.d.). *Epidermis (E), Dermis (D), Basal layer (B), Hair follicle (HF) **All counted cells are DAPI positive ***White scale bar, 50 µm