Supplementary Material
Supplementary Figure 1. KY19382 elevates Wnt/β-catenin signaling, hair induction markers, and proliferation. (a) Rat DP cells were treated with the indicated concentration of KY19382 for 48 hours. Cell viability was measured in rat DP cells that were treated with the indicated concentrations of KY19382 in 24 wells. (b) Rat DP cells were treated with the indicated concentrations of KY19382 or DMSO for 48 hours. Immunoblotting was performed to detect β-catenin and PCNA. (c) Immunocytochemical analysis for β-catenin (green) of rat DP cells. (d) Rat DP cells treated with 1 or 5 µM of KY19382 were subjected to ALP staining via incubation with BCIP/NBT. Arrow indicated ALP-positive rat DP cells. (e) The ALP activity was quantified and normalized with respect to total protein concentration. Scale bars = 50 µm. Values are expressed as means±SEM. Student’s t-tests (* P < 0.05, ** P < 0.01, *** P < 0.001, n=6).
Supplementary Figure 2. Human DP cells transfected with control siRNA or β-catenin siRNA. Human DP cells were treated with 0 or 5 μM KY19382. Transfection of β-catenin siRNA was confirmed through immunoblotting.
Supplementary Figure 3. KY19382 accelerates rat vibrissa follicle elongation. Rat vibrissa follicles from Wistar were cultured with 1 or 5 μM KY19382 in DMEM for 12 days. (a) Elongation rate of rat vibrissa follicles was calculated as the difference in the length of vibrissa follicles wherein vibrissa follicle length in the control group at day 12 was considered 100% (n=8). (b) IHC analysis of rat vibrissa follicle hair bulb for β-catenin (green). Dashed lines were rat vibrissa follicles. (c) IHC analysis of rat vibrissa follicle bulge for β-catenin. Arrows indicate β-catenin positive nuclei. Scale bars = 50 µm. Student’s t-tests (* P < 0.05, ** P < 0.01, *** P < 0.001).
Supplementary Figure 4. Treatment with KY19382 increases proliferation marker in the dorsal skins of mice. Mouse dorsal skins were treated for 14 days. IHC staining for PCNA. Scale bars = 100 µm.
Supplementary Figure 5. Treatment with KY19382 enhances proliferation marker in mice wounds. Mice wounds were treated for 14 days. IHC analysis for PCNA. Scale bars = 100 µm.