Hair plucking is the most frequently used method of anagen induction within hair follicles. In this study, we found that plucking leads to the entire renewal of the follicular stem cell region of the mouse pelage follicle. Comparative histochemical analysis revealed that S100A4 protein was specifically distributed in the outer layer of the epithelial sac, which has been identified as the stem cell region of the pelage follicle, whereas the slow cycling cells that retained 5-bromo-2'-deoxyuridine label for 8 wk were located in the epithelial sac and also in the hair germ. Combined terminal deoxynucleotide transferase deoxyuridine triphosphate fluorescein nick end labeling method and immunohistochemistry revealed that positive cells were detected in the outer layer of the epithelial sac possessing both bromo-2'-deoxyuridine and S100A4 labels 4.5 h after plucking. No terminal deoxynucleotide transferase deoxyuridine triphosphate fluorescein nick end labeling signal, however, was observed in the hair germ. Serial inspection of the plucked follicle revealed that almost all regions of the epithelial sac became terminal deoxynucleotide transferase deoxyuridine triphosphate fluorescein nick end labeling positive 12 h after plucking. Terminal deoxynucleotide transferase deoxyuridine triphosphate fluorescein nick end labeling-positive cells ultimately degenerated without forming apoptotic bodies. Subsequently, the surviving label-retaining cells in the hair germ migrated upward to re-epithelialize the damaged portion. These results indicate that follicular stem cells in the epithelial sac underwent cell death after plucking. It is likely that the hair germ is responsible for the reconstruction of the stem cell region of the hair follicle.