FIGURE 3.

KY19382 stimulates hair regrowth in vivo. C57BL/6N mice were treated with Vehicle 1 (PEG400), Vehicle 2 (50% [v/v] ethanol, 30% water and 20% propylene glycol), 0.5‐mM KY19382 or 100‐mM minoxidil (MNX) for 14 or 28 days (n = 5). (a) The gross image showed hair regrowth in mice treated with each drug for 28 days. (b) Quantitative measurements of the weight of regrown hairs. (c) Haematoxylin and eosin (H&E) staining to evaluate the hair follicles of skins with different treatments. (d, e) Quantitative analyses of the relative number of hair follicles and dermal thickness of H&E staining images (n = 5). (f) Immunohistochemistry (IHC) staining for keratin 15, β‐catenin and Ki67 using the dorsal skin of mice treated with each drug for 14 days. Lines show keratin 15‐positive bulge stem cells region. (g, h) Quantitative estimations of the total and nuclear β‐catenin intensities in bulge stem cells (n = 15 taken in five different images). (i) Alkaline phosphatase (ALP) staining was performed to evaluate the ALP activity of hair follicles with different treatments. Arrow points to the ALP‐positive region. (j) Immunoblotting analyses for β‐catenin, PCNA and ERK. Scale bars = 100 μm. Values are expressed as means ± SEM. * P < .05, significantly different as indicated ; NS, not significant