Total RNA was isolated from kidneys of these mice, and mRNA was quantified by the branched DNA signal amplification assay. The data is plotted as Trichostatin A mouse average RLU per 10 μg total RNA ± SEM. B) Protein expression of Slco1a1 and 1b2 in crude membrane fractions from kidneys of C57BKS and db/db mice (n = 2). Proteins (75 μg/lane) were separated on 4–20% acrylamide/bis PAGE, transblotted, incubated with primary and secondary antibodies and visualized by fluorescence. C) Quantification of western blots by

using the Quantity One® software (Biorad, Hercules, CA). The average band intensity for C57BKS males was considered 100% and other groups were compared with that density. Alvocidib cost Asterisks (*) represent a statistically significant expression difference between db/db mice and C57BKS control mice of the same gender (p≤0.05). Number signs (#) represent a statistically significant expression difference between male and female db/db mice or male and female C57BKS mice (p≤0.05). Slc22a7 mRNA expression was downregulated in db/db male and female mice. Slco1a1, Slc22a2 and 22a6 mRNA expression INCB018424 solubility dmso was downregulated in db/db males as compared to C57BKS males. Slco1a1, Slc22a2 and 22a6 mRNA expression was more in C57BKS males as compared to C57BKS females. Slco1a1 and 1b2 protein expressions were significantly decreased in db/db females as compared to C57BKS females. Figure 6 Efflux transporter expression in kidneys of C57BKS and db/db mice. A) Messenger RNA expression

of Abcc3, 4 and Abcb1. Total RNA was isolated from kidneys of adult db/db and C57BKS mice, and mRNA expression was quantified using the branched DNA signal amplification assay. The data plotted as average RLU per 10 μg total RNA ± SEM. B) Protein expression of Abcc4 from crude membrane fractions of kidneys of C57BKS and db/db mice (n = 2). Proteins (75 μg/lane) were separated on 4–20% acrylamide/bis PAGE, transblotted, incubated with primary and secondary antibodies and visualized by fluorescence. C) Quantification of western blot by using the Quantity One® software (Biorad, Hercules,

CA). The average band intensity for C57BKS males was considered Palmatine 100% and other groups were compared with that density. Asterisks (*) represent a statistically significant expression difference between db/db mice and C57BKS mice of the same gender (p≤0.05). Number signs (#) represent a statistically significant expression difference between male and female db/db mice or male and female C57BKS mice. Abcc3 expression was downregulated in db/db females and upregulated in db/db males as compared to respective controls. Abcc4 mRNA expression was upregulated in db/db males as compared to C57BKS males. Abcc1, 2, Abcg2 mRNA expression also remained unchanged in kidneys of these mice (data not shown). Among efflux transporters, expression of Abccs was altered in kidneys of db/db mice. Db/db females exhibited marked down regulation of Abcc3 mRNA in kidney compared to C57BKS female mice.