Figure 3

Full-length Klotho mRNA is upregulated in CMP hearts. (A) 3′RACE PCR strategy utilizing oligo dT primers to detect the full-length and secreted isoform of Klotho mRNA in the human heart. (B) Columns 1–2 (Human Tissue): RT-PCR expression analyses of the full-length (fl-Klotho) and the secreted form (s-Klotho) of Klotho mRNA in human CMP hearts (H) and kidney (K) as positive control. Beta actin was used as internal control. Columns 3–6 (Mouse Tissue): RT-PCR expression analyses of Klotho in KO H (−/−) and WT H (+/+) hearts as well as Klotho KO K (−/−) and WT K (+/+) kidney as controls. The gels were cropped for conciseness. Full length gels are presented in Supplementary Figure S3 and S4. (C) Relative amount of Klotho mRNA levels related to the reference gene RPL32 in human kidney (n = 4), control hearts (n = 10) and CMP hearts (n = 10). Shown are all individual data points, lines show mean ± SD. ^**P ≤ 0.01. (D) Relative amount of FGF23 mRNA levels related to the reference gene RPL32 in human kidney (n = 4), controls hearts (n = 10) and CMP hearts (n = 10). Shown are all individual data points, lines show mean ± SD. ^***P ≤ 0.05. FGF mRNA was not detected (ND) in healthy kidneys.