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. 2013 Jan;41(1):170–179. doi: 10.1124/dmd.112.049171

Fig. 5.

Fig. 5.

Effect of UGT2A2_i2 coexpression on UGT2A2_i1 activity against PAH substrates. (A) a representative RT-PCR gel showing UGT2A2Δexon3 coexpression with wild-type (WT) UGT2A2 following transient transfection into HEK293 cells stably overexpressing UGT2A2_i1. No UGT2A2Δexon3 transcript was detected using RNA from the untransfected cells. The cDNA equivalent of 100-ng RNA was used for each RT-PCR reaction. (B) real-time PCR was completed using the assay described in Fig. 1 to determine relative wild-type UGT2A2 and UGT2A2Δexon3 levels following transient transfection. cDNA corresponding to 20 ng of RNA was used in each reaction, with levels of wild-type UGT2A2 and UGT2A2Δexon3 expression determined relative to wild-type UGT2A2 expression in the untransfected cells. As expected, only trace amounts of UGT2A2Δexon3 were detected in the untransfected UGT2A2_i1 overexpressing cells. (C) effect of UGT2A2_i2 coexpression on UGT2A2_i1 activity against 1-OH-pyrene and 8-OH-B(a)P using a substrate concentration at the previously determined KM for UGT2A2. Glucuronidation rates were corrected to account for differences in the relative level of UGT2A2_i1 expression in the untransfected cells and cells transiently transfected with UGT2A2_i2. AU, arbitrary unit.