Fig. 2: Expression of hair follicle lineage markers by transplanted bladder epithelial cells.

a Microscopic appearance of a hair follicle bulb (left) and of a sebaceous gland (right) generated by epithelial cells cultured from the bladder of an EGFP rat and transplanted into a newborn mouse skin microenvironment (103 and 238 days after transplantation, respectively), n = 3 mice. EGFP expression (green) was revealed by immunohistochemistry, nuclei (blue) were counterstained with Hoechst 33342. Note that the transplanted EGFP cells contributed to all the differentiated lineages of the hair follicle and sebaceous glands; bars: 100 μm. b Expression of several transcription factors important for hair follicle differentiation as determined by qPCR analysis. ΔCt results were normalized against housekeeping genes (TBP, SDHA, and Tubb). Mean of 3–9 technical replicates with individual data represented by empty circles. c Transplanted EGFP-positive bladder epithelial cells expressed SOX9, LHX2, cytokeratin-15 (CK-15), and cytokeratin-31 (CK-31) at the proper location in the hair follicles. For each marker, n = 3 experiments. Hair follicles and sebaceous glands generated from multipotent epithelial stem cells cultured from a whisker follicle were used as control. SOX9 (red): whisker: day 103 after transplantation, bladder: day 103; LHX2 (red): whisker: day 238, bladder: day 236; CK-15 (red): whisker: day 103, bladder: day 238; CK-31 (red): whisker: day 236, bladder: day 103. EGFP expression (green) was revealed by immunohistochemistry, nuclei (blue) were counterstained with Hoechst 33342. Bars: 100 μm.