Fig. 2

SOX11 overexpression inhibits epidermal differentiation. a Constructs used to generate transgenic mice expressing epithelial-specific Sox11 under the control of tetracycline and its derivatives. b Immunofluorescence analysis of expression of FLAG-tagged SOX11. Four-day old K14-rtTA;TRE-Sox11-FLAG mice were injected intraperitoneally with Dox for the indicated time prior to skin isolation. Dashed line demarks the border between the epidermis and dermis. c Images of embryos of specified genotypes in X-gal exclusion assay. The pregnant females were on Dox-containing diet until the embryos were collected at E18.5. d Immunofluorescence analysis of differentiation markers in postnatal epidermis expressing SOX11. After birth, the mother was on Dox-containing diet for 5 days and skin sections from K14-rtTA (control) and K14-rtTA;TRE-Sox11-FLAG pups were immunostained with the indicated antibodies. KRT5 (keratin5); KRT1 (keratin 1); LOR (loricrin); FLG (filaggrin). e Immunochemical analysis of embryonic makers in control and SOX11-FLAG expressing epidermis with the indicated antibodies. KRT18 (keratin 18); TCF7L1 (transcription factor 7 like 1); TCF7L2 (transcription factor 7 like 2). (f–j) Effect of long-term SOX11 overexpression in postnatal skin. Dorsal skins from K14-rtTA (control) and K14-rtTA;TRE-Sox11-FLAG newborns were grafted in pair onto Nude mice, which were put on a diet containing Dox day 10 post grafting. n = 2. f Image of engrafted mouse 28-day post grafting with engrafted area demarcated by yellow dashed lines. g H&E staining of the sectioned pair of skins showing stunted hair follicles (*) in SOX11-induced skin. f Oil red O staining marks lipids, enriched in sebaceous glands. Immunofluorescence analysis of differentiation markers LOR (i) and FIL (j) in grafted skin. White dashed lines mark the border between epidermis and dermis. HF, hair follicles; Es, eschar; SG, sebaceous glands. Scale bars, 50 µm (b, e) 20 µm (d, i–j), 100 µm (g, h). All images reflect the same result seen across the entire embedded skin section from 2 (b, c and f–j) or 3 (d and e) biological replicates