FIGURE 5.

A, analysis of Ugt3a1 and Ugt3a2 mRNA levels in neonatal mouse (m) liver and kidney by quantitative RT-PCR. Data were analyzed using a standard curve for each gene and normalized to the housekeeping gene Rps26. B, analysis of the enzymatic activity of mouse Ugt3a2 with its preferred aglycone substrate hyodeoxycholic acid (HDCA) and either UDP-Glc or UDP-GlcNAc as the sugar donor. Assays were chromatographed and visualized by autoradiography. The arrow indicates the hyodeoxycholic acid glucoside, and the asterisks indicate endogenous glycosidation activities in HEK293T cells. n.c., negative control HEK293T lysate. C, analysis of endogenous glucosidation and N-acetylglucosaminidation activities in mouse liver and kidney microsomal preparations. Assays were performed using a range of aglycone substrates and either UDP-Glc or UDP-GlcNAc as the sugar donor and were analyzed as described for B. MCA, mycophenolic acid; 4-MU, 4-methylumbelliferone; UDCA, ursodeoxycholic acid.