Extended Data Fig. 9: Analyses of skin changes upon Gas6 overexpression or treatment with an AXL inhibitor.
From: Corticosterone inhibits GAS6 to govern hair follicle stem-cell quiescence
a, Immunohistochemical analysis (GFP and PCAD) of PBS-injected second telogen skin and AAV-GFP-injected second telogen skin. Dashed lines, epidermis and hair follicles; solid lines: DP. b, RT–qPCR of Gas6 from dermal fibroblasts of PBS-injected second telogen skin (control) and AAV-CAG-Gas6-injected second telogen skin. c, Precocious HFSC activation in mice injected with AAV-CAG-Gas6 shown by EdU incorporation. Immunocolocalization (EdU and CD34) in control and AAV-CAG-Gas6-injected skin after AAV injection (D3 to D9). d, Comparison of EdU and CD34 localization in bulge and upper outer root sheath (ORS) in late anagen (AnaV) (control, D50 after injection; GAS6, D17 after injection). e, H&E staining of late anagen (AnaVI) skin (control, D53 after injection; GAS6, D20 after injection). Quantification of the ORS length in the zigzag hairs of control and AAV-CAG-Gas6-injected mice during late anagen. Brackets indicate the ORS length below the bulge. f, Immunocolocalization (EdU and CD34) in infundibulum, junctional zone, sebaceous gland, mid ORS, lower ORS and matrix of late anagen (AnaVI) hair follicles in control and AAV-CAG-Gas6-injected mice with quantifications. Dashed lines outline hair follicles. g, Top, H&E staining in the late anagen skin of control and AAV-CAG-Gas6-injected mice with quantification of the thickness of epidermis (E). Bottom, immunocolocalization (EdU and DAPI) in interfollicular epidermis and dermis in control and AAV-CAG-Gas6-injected mice with quantifications. h, RT–qPCR of genes related to HFSC proliferation in HFSCs of second telogen skin. i, Hair cycle progression of sham and ADX mice treated with ethanol topically, or ADX mice treated with R428 in ethanol. j, RT–qPCR of genes related to cell-cycle machineries and cytokinesis from HFSCs of sham and ADX mice treated with ethanol topically, or ADX mice treated for 7 days with R428 (in ethanol) topically. Yellow dashed lines, bulge (c, d); white dashed lines, hair germ (c), the rest of the hair follicle (d, f), or the boundary of epidermis and dermis (g); solid white line, DP (c). EtOH, ethanol. Scale bars (a, c–g), 50 μm. Data are mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS, not significant. For exact P values, see Source Data. For statistics, sample sizes and numbers of replications, see Methods.
