Figure 5.

Nestin. (A) RT-qPCR confirmed the presence of Nestin mRNA in comparison to 18S rRNA; n=10. (B) Representative western blots. Left panel: Recombinant truncated V5-tagged Nestin was expressed in E. coli and lysates probed on the same membrane with indicated antibodies. The negative control samples were the same as that in Fig. 1. Anti-V5 and AHP1739 anti-Nestin (AbD Serotec) antibodies detected a band at 25 kDa in size. The expected size of the recombinant truncated Nestin protein is 22 kDa. Right panel: Dog skin lysates probed with N1602 anti-Nestin antibody (IBL) revealed a band above 250 kDa and two nonspecific bands of 55 and 75 kDa. Procedures were as per those specified in Fig. 1. (C) Schematic drawing and immunohistochemical staining for Nestin. Picture on the left: Anagen HF with scattered, positively stained cells within the dermal papilla. Picture in the middle: Magnifications of dermal papillae. Note the cytoplasmic staining of the dermal papilla and the connective tissue sheath. Picture on the right: Telogen HF. The cells did not show any positive staining in the entire HF; n=5. Note that, for a better overview, the schematic drawings of the hair follicles depict only one follicle of each cycle stage in which a positive staining was observed. Scale (C, left) 100 µm; (C, middle) 25 µm; (C, right) 50 µm.