Fig. 5.

Uptake transporters Slco1a1 and 1a6, Oat1 and 2, and Oct1 and 2 expression in kidneys of lean control and DIO mice. A, mRNA expression for uptake transporters organic anion-transporting polypeptide Slco1a1 and 1a6 and organic anion and cation transporters Oat1 and 2 and Oct1 and 2 in kidneys of lean control and DIO mice. Total RNA was isolated from kidneys of mice by phenol chloroform extraction, and mRNA was quantified using a branched DNA signal amplification assay. The data are plotted as average relative light units (RLU) per 10 μg of total RNA ± S.E.M. B, protein identification and quantification by Western blotting for Slco1a1 in the crude membrane fraction from kidneys of lean control and DIO mice. The proteins were isolated by electrophoresis on 10% acrylamide/bis gels, transblotted, incubated with primary and secondary antibodies and substrate, and detected by chemiluminescence. C, quantification of Western blots with Quantity One software. The average band intensity for lean mice was considered to be 100%, and band intensities in the other groups were compared with it to plot the graphs. *, statistically significant difference of expression between lean control and DIO mice (p ≤ 0.05). Only Oat2 mRNA expression was down-regulated in DIO mice; Slco1a1 and 1a6, Oat1, and Oct1 and 2 mRNA remained unchanged compared with that in lean mice. Slco1a1 protein expression also remained unchanged. Gapdh, glyceraldehyde-3-phosphate dehydrogenase.