Fig. 2.
Molecular analyses of anagen HFs from WT and TG mice suggest differences in expression of cell-cycle-associated genes. Skin of 12-week-old WT and TG mice was synchronized at anagen VI stage of the hair cycle by depilation. Skin was harvested, and cryosections were processed for laser-capture microdissection, RNA isolation and amplification, microarray and real-time PCR analyses (A–G), and for immunohistochemistry (H–J). (A and B) Anagen VI HF before and after laser-capture microdissection. (C) RT-PCR of the hair matrix- and FP-specific genes in RNA samples obtained from the corresponding areas of anagen VI HFs of WT and TG mice. (D–G) Real-time PCR of genes expressed in the hair matrix or FP that show differences between intermediate HFs of TG mice and intermediate or small HFs in WT mice. (H) pSmad1/5 in the hair matrix (large arrows), hair shaft (small arrows), and outer root sheath (arrowheads) in small and intermediate HFs of WT and TG mice. (I) Ectopic appearance of BrdU+ (green fluorescence, arrowhead) and Ki-67+ cells (red fluorescence, arrowhead) in differentiating cells of the hair shaft in TG mice. (J) Decrease in p27Kip1 expression in hair matrix of intermediate HFs in WT and TG mice (arrows).