Figure 3.

Keratin15. (A) RT-qPCR confirmed the presence of K15 mRNA in comparison to 18S rRNA; n= 10. (B) Representative western blots. Left panel: Recombinant truncated V5-tagged K15 was expressed in E. coli and lysates probed on the same blot with indicated antibodies. The negative control samples were the same as in Fig. 1. Anti-V5 and anti-K15 antibodies detected the expected band of 39 kDa in size. Right panel: Dog skin lysates probed with the same anti-K15 antibody revealed a single band at the expected size of canine K15 of 55 kDa, which was also weakly detected with the secondary antibody alone. Procedures were as per those specified in Fig. 1. (C) Schematic drawing and immunohistochemical staining for K15. Picture on the left: Anagen HF with positive staining in the ORS basal cells of the upper isthmus. Picture in the middle: Magnification of the picture on the left. Note positive cytoplasmic staining. Picture on the right: Telogen HF with positive staining in the ORS of the lower half of the isthmus; 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.