Figure 3: Changes in expression of Klotho in tubular cells (MDCK cells) after exposure to high concentrations of glucose. Cultured MDCK cells exposed to glucose at final concentrations of 10 mM, 20 mM, 30 mM and 40 mM were used to compare with the control, which was incubated with 5.5 mM glucose. Cells were also exposed to 24.5 mM mannitol to produce the same osmolarity (317 mOsmol/L) as that produced when using the highest concentration of glucose (30 mM). After 24 hours of incubation, cells were prepared for Western blot analysis. Quantification of protein levels using Klotho over 𝛽 -actin indicated as means with SE ( 𝑛 = 4 per group) in each column are presented in the lower panel. 𝑃 < . 0 5 , 𝑃 < . 0 0 1 compared to control.