JP-2025516076-A5 -

Dates

Publication Date

20260511

Application Date

20230518

Description

The images illustrate OR2A4/7 mRNA expression in the olfactory epithelium (upper panel), hair follicle (HF) hair bulb (center panel), and bulge (lower panel) (indicated by dark gray pink arrows in the black and white version of the diagram).Representative images of OR2A4/7 protein expression in olfactory epithelium (positive control, upper panel) and fresh human scalp epidermis (lower panel) are shown as examples.Examples of various HF compartments in fresh human scalp (n=3, independent donors) are shown. CTS: connective tissue sheath, DP: dermal papilla, IRS: inner root sheath, ORS: outer root sheathThe image (left panel) shows OR2A4/7 protein expression in various compartments of organ cultured HF (ex vivo) hair bulbs (green, light gray in the black and white version of the figure), and the bar graph (right panel) shows a quantitative analysis of pooled data for OR2A4/7 expression in the dermal papilla, hair matrix (germ + precortical), and outer root sheath under treatment with vehicle or 50 μM CHS.The bar graph (left panel) shows a quantitative analysis of pooled data for OR2A4/7 expression in the basal and upper basal layers of the bulge under treatment with vehicle or 50 μM CHS, and the representative image (right panel) shows the bulge from HF treated with vehicle and 50 μM CHS, exhibiting OR2A4/7 (green, white in the black and white version of the figure) and integrin α-6 (red, light gray in the black and white version of the figure). The nuclei are counterstained with DAPI (blue, dark gray in the black and white version of the figure). All data are presented normalized to a control group, each treated with vehicle (mean ± SEM, n=28–49 HFs from n=5–8 independent donors, *p<0.05, ***p<0.001).An example bar graph shows the quantitative analysis of K15+ stem cells in the basal layer of the bulge under vehicle or 50 μM CHS treatment.The image on the left shows a representative image of CD34 protein expression in ex vivo HF (red, light gray in the black and white version of the figure), and the bar graph on the right shows the quantitative analysis of CD34+ cells in the basal layer of the outer root sheath (ORS) under vehicle or 50 μM CHS treatment; no difference is observed.The images show a representative image (left) of CD71 (red, light gray in the black and white version) and OR2A4/7 (green, white in the black and white version) protein co-expression in ex vivo HF, a bar graph (middle) showing quantitative analysis of CD71+ cells in the hair matrix and proximal ORS, and a bar graph (right) showing the percentage of CD71/OR2A4/7 double-positive cells in the proximal outer root sheath. The nuclei are counterstained with DAPI (blue, dark gray in the black and white version). All data are normalized to a control group, each treated with the vehicle (mean ± SEM, n=36–44 HF cells from n=8 independent donors, *p<0.05, *p<0.01, ***p<0.001).The percentage of HF in each hair cycle under culture with vehicle or 50 μM CHS is illustrated. Green arrows (gray in the black and white version of the figure) indicate the increase in HF during the growth phase under 50 μM CHS treatment. n=8 independent donors.The left panel shows representative images of Ki- 67 (red, light gray in the black and white version) and TUNEL (green) protein expression in ex vivo HF hair follicles for analyzing cell proliferation; the right panel shows representative images of Masson-Fontana staining in ex vivo HF hair follicles for melanin staining; and a bar graph shows the quantitative analysis of Ki-67 protein expression in hair matrix keratinocytes under vehicle or 50 μM CHS treatment. The nuclei are counterstained with DAPI (blue, dark gray in the black and white version). All data are presented normalized to a control group treated with the vehicle (mean ± SEM, n=45–47 HF cells from n=8 independent donors, *p < 0.05, ***p < 0.001). OR2A4/7 stimulation also prolongs the growth phase (Figure 4a) and induces hair matrix keratinocyte proliferation in ex vivo HF (Figure 4b).