Figure 2

Expression profiles of Klotho in intervertebral discs. (A) Sagittal sections from 3-week-old (a-d, i) and 12-week-old (e-h, j) rats. Sections were treated with anti-Klotho antibody and counterstained with hematoxylin. Note that nucleus pulposus cells expressed Klotho protein (arrows in d and h). A magnified view of the areas (panel b and f) is marked by the square in panels c, d, g, and h. (B) Real-time RT-PCR analysis of Klotho mRNA levels in nucleus pulposus (NP) and annulus fibrosus (AF). Scale bar, 50 μm to 1.0 mm (original magnification, × 4, × 20, × 40). (C) Immunofluorescence was performed after treatment of BIO in nucleus pulposus cells. The subcellular distribution of β-catenin is shown in red; Klotho, in green; and degree of merge, in yellow.