DMD #10520

Table s1. Oligonucleotide probes generated for analysis of mouse PXR, Mdrs, Bsep, and Ibat mRNAs expression by Quantigene branched DNA signal amplification assay

Functiona / Probe Sequence / Function / Probe Sequence
Bsep (NM_021022b) / Ibat (Cont’d)
CEc / ggtcgagaaggatatgtaaatttacaTTTTTctcttggaaagaaagt / LE / tttgtctcatcaaatgatggccTTTTTaggcataggacccgtgtct
CE / atgtttgaacggaggaactgaaTTTTTctcttggaaagaaagt / BL / gttcaatgatccaggcactttg
CE / tttgcggcagctatagctctcTTTTTctcttggaaagaaagt / BL / agtctagccaggaagaaaccca
CE / ccgcgagagagttgagagccTTTTTctcttggaaagaaagt / BL / gctgagtgttctgcattccagttt
CE / ccctctctggctttgtccagagTTTTTctcttggaaagaaagt / BL / atcctcgggggagaaggag
LE / tgcccaggatccacagataccTTTTTaggcataggacccgtgtct / BL / ggaaggtgaacaccaggttgag
LE / ccaacgaatgccagcgtcTTTTTaggcataggacccgtgtct / BL / gagctggaaaacagtatagatgagtg
LE / gcttttaccacacccactgctcTTTTTaggcataggacccgtgtct / BL / attcctaatattactgctgcaaagac
LE / tccaacagctggatgctggtTTTTTaggcataggacccgtgtct / BL / tctctagaaactcagcatcatttttt
LE / tgatcgggatcgtagaaccgtTTTTTaggcataggacccgtgtct / BL / tctcatctggttgaaatcccttg
LE / ccatctatcatcaccgttcccTTTTTaggcataggacccgtgtct / BL / gaagttttctttttctttcttgtctact
LE / cattgacttttttgctgtcgtgaTTTTTaggcataggacccgtgtct / BL / tgattttccaccaaaagaaataaa
LE / gctcctgggagacaatcccaTTTTTaggcataggacccgtgtct / BL / tttatacaaaaaacataatttcaatttga
LE / cattatgctacagtcaaataacacggTTTTTaggcataggacccgtgtct / BL / tctttgtcacactaaagaatggattac
LE / ttgtccccatacttgatgttgtcTTTTTaggcataggacccgtgtct / Mdr1a (NM_011076)
LE / agtcatgcagctgagcctgcTTTTTaggcataggacccgtgtct / CE / gcattttaaagcattaatctccattTTTTTctcttggaaagaaagt
LE / ctggatcccaacattagtttcatatTTTTTaggcataggacccgtgtct / CE / ctcaggtgcccatgtctgagtTTTTTctcttggaaagaaagt
LE / aatagcaatgcgttgtttctccTTTTTaggcataggacccgtgtct / CE / tgagcctgcaggatctgtctttTTTTTctcttggaaagaaagt
LE / tcgtacaatggcccgagcTTTTTaggcataggacccgtgtct / CE / catttgcttttccctctgattttTTTTTctcttggaaagaaagt
LE / catccagtagcaagattttaggatcTTTTTaggcataggacccgtgtct / LE / ccaaactaaatgctttccactgtgTTTTTaggcataggacccgtgtct
LE / tctgtgtctaaggcagatgtagcttTTTTTaggcataggacccgtgtct / LE / tgatgctgtatctatttttgtttttttaTTTTTaggcataggacccgtgtct
LE / caagctgcactgtcttttcacttTTTTTaggcataggacccgtgtct / LE / cacttttaacccttttataaattttatagtTTTTTaggcataggacccgtgtct
LE / agcaatgacaatacaggtccgaTTTTTaggcataggacccgtgtct / LE / cagtaatttttgatacatacataatttgttaTTTTTaggcataggacccgtgtct
LE / agttctggatagtagacaagcgatgTTTTTaggcataggacccgtgtct / LE / cagtttcactatatatactttatacaaacatttTTTTTaggcataggacccgtgtct
LE / gacatgacggcgatgatatctgTTTTTaggcataggacccgtgtct / LE / aatatataatttcgcatgcatttgaTTTTTaggcataggacccgtgtct
BL / gacagtccattcagaacttgtatatca / LE / ttatgtacaatatcccagttgggacTTTTTaggcataggacccgtgtct
BL / tccacggagatctctttggtg / LE / cttacttaggtggtagggatctgaaTTTTTaggcataggacccgtgtct
BL / ttctctgggagtgacatgacga / LE / acaaaaacaccacaccttgtctgTTTTTaggcataggacccgtgtct
Ibat (AB002693) / LE / tctgcctctagcactggcattTTTTTaggcataggacccgtgtct
CE / ggctgtagccagctatagggaatTTTTTctcttggaaagaaagt / LE / ggctgtttggccagccacTTTTTaggcataggacccgtgtct
CE / ttcagttttagtttaatgaatattgttctTTTTTctcttggaaagaaagt / LE / ctgcatgtatggcatttgcatTTTTTaggcataggacccgtgtct
CE / atttaacttttaaatttgttttttctgaTTTTTctcttggaaagaaagt / LE / tgtgcgtgtgtgtgcacatacttTTTTTaggcataggacccgtgtct
LE / attgttcctataatccacagtttggTTTTTaggcataggacccgtgtct / BL / attacaaagcctctgaagactctaaa
LE / cctgtaccagggttgaccagctTTTTTaggcataggacccgtgtct / BL / gaaagtataaataaaagtatatattatcaaacatt
LE / ccaaggctactgttcggcaTTTTTaggcataggacccgtgtct / BL / agcagaaatcagttatagttacaaatga
LE / agctgtacaatggtggagcacaTTTTTaggcataggacccgtgtct / BL / gcaaaataactcaaaaatatgaatgct
LE / ccataacatttcctgtatgtgacataaTTTTTaggcataggacccgtgtct / BL / tggactgttttttaaaggctaaaa
LE / tggagtccatttcattgtctgtttTTTTTaggcataggacccgtgtct / BL / ccccctccccccagtaa
Mdr1a (Cont’d) / Mdr2 (Cont’d)
BL / aacactttgcccatagagtgatg / BL / cggctggagggctccatt
BL / ggaacgcgctatgttggcta / BL / tcagacaggatgatggagaaagc
BL / tgagacaagttttctcactgaacct / BL / cagggccaaagatagctatcatc
BL / gagtgcacatggctgacacct / BL / actcctaggcccaagaagacc
Mdr1b (NM_011075) / BL / aaggaagaaagtaaagaaggagagg
CE / ccttattaaaggtgatttggattctTTTTTctcttggaaagaaagt / BL / gtggtgaggatctctccagctt
CE / ttagatttagatttaggatccgccTTTTTctcttggaaagaaagt / BL / gccatggaccggagcctt
CE / cagcccttgccttttaactttTTTTTctcttggaaagaaagt / BL / gtagaaagtgctccagtactgttttta
LE / attatttcctggttcaatttcatttTTTTTaggcataggacccgtgtct / BL / cgtggctccttggacttgc
LE / tgtcactctgggatccataagcTTTTTaggcataggacccgtgtct / BL / caattaaagccaacttggttcc
LE / ctgtggacacttctgtaaattgatctTTTTTaggcataggacccgtgtct / BL / cagcagaagtgtcagttgccaa
LE / caacaagtaaataaggccattcacTTTTTaggcataggacccgtgtct / BL / gcaatgaatggaacaaccgataa
LE / aaacactggttgtatgcacccaTTTTTaggcataggacccgtgtct / BL / gatacaacagttcgaatattttctattg
LE / gctccagcctggaccattgTTTTTaggcataggacccgtgtct / BL / tcaaattttctttcttgggtcaag
LE / gtcacagctcatgagcgctttTTTTTaggcataggacccgtgtct / BL / atgcaatttttcaacatacattgac
LE / caatattaaaaatacttagcacctcagataTTTTTaggcataggacccgtgtct / Murat1 (NM_009203)
LE / ggtttggtgccatgtttaaacacTTTTTaggcataggacccgtgtct / CE / tcgcctgggaggtgctgtTTTTTctcttggaaagaaagt
LE / tttgtttccgattttggttcatTTTTTaggcataggacccgtgtct / CE / ctctgtaagctgccattgaggtTTTTTctcttggaaagaaagt
LE / ctatggcttgtttgtttgtttgttTTTTTaggcataggacccgtgtct / CE / caggaagatggactgggccTTTTTctcttggaaagaaagt
LE / attaaaaacatggccctgtttaaTTTTTaggcataggacccgtgtct / LE / tgtccaggagaggcacccaTTTTTaggcataggacccgtgtct
BL / tctgaagtcagttcagaagcatcag / LE / tccaaggtccccagggatacTTTTTaggcataggacccgtgtct
BL / aagtcttctctcttggtcttgcttt / LE / ggaccaggtgggatggagaTTTTTaggcataggacccgtgtct
BL / ttcatccacagcctctttcatact / LE / ggcagcgtcactccagttggTTTTTaggcataggacccgtgtct
BL / aaaaggaaaccagaggcacatc / LE / ccatcctcgcatggctcagtTTTTTaggcataggacccgtgtct
BL / tttataacagcgcaaagtacgc / LE / ggtgctatggtcgtaaacccagTTTTTaggcataggacccgtgtct
BL / tctcatcttgatggagttacctttaa / LE / gagttacataccaggtcccacgtTTTTTaggcataggacccgtgtct
BL / ttaattacaaagtctcggaaggct / LE / atgggtctcagggcctggTTTTTaggcataggacccgtgtct
Mdr2 (NM_008830) / LE / ggctcctaccaggattccagcTTTTTaggcataggacccgtgtct
CE / tcccaaacgtgaagccctgTTTTTctcttggaaagaaagt / LE / ggcatggccacacactgcTTTTTaggcataggacccgtgtct
CE / cgtaaataaatgatataataataccggtTTTTTctcttggaaagaaagt / LE / tgcgcccaaacctatctgaTTTTTaggcataggacccgtgtct
CE / gcttccatttcctttttatctctctTTTTTctcttggaaagaaagt / BL / cggccagaagaacatcggg
CE / cctctgttgcaatctttccagcTTTTTctcttggaaagaaagt / BL / tggtggggttgctggtca
CE / gcaccgaattcctgtaaggtccTTTTTctcttggaaagaaagt / BL / tgtcggaagcggaggcac
LE / ggcaatggcacagactgttccTTTTTaggcataggacccgtgtct / BL / tggccgtggcgttgga
LE / tgcttcactgcgtcatcccTTTTTaggcataggacccgtgtct / BL / ggtcacgattgtggacctgaa
LE / agggagaacatgttacacttttgcTTTTTaggcataggacccgtgtct / Npc1L1 (NM_207242)
LE / ctgccttagcatcgctttaaagTTTTTaggcataggacccgtgtct / CE / cctagaaagaagcagatggcctTTTTTctcttggaaagaaagt
LE / tgatcatcaaaccagctcatgtcTTTTTaggcataggacccgtgtct / CE / agatgattgctaagccagaggtcTTTTTctcttggaaagaaagt
LE / gcagcatctgtggcgagtcttTTTTTaggcataggacccgtgtct / CE / tgcctcttgctatccagggagTTTTTctcttggaaagaaagt
LE / ccaaggttggctgtgttctgtgTTTTTaggcataggacccgtgtct / CE / tgcagcaggaagggagtgtatatTTTTTctcttggaaagaaagt
LE / ttcatttcaacaattcctgctacaTTTTTaggcataggacccgtgtct / LE / cagagaggctgcacagcagcTTTTTaggcataggacccgtgtct
LE / ggcattgccagccaacattTTTTTaggcataggacccgtgtct / LE / ggcatggaggtcagggccTTTTTaggcataggacccgtgtct
Npc1L1 (Cont’d) / PXR (Cont’d)
LE / aaggcaaaggtcctcacagctTTTTTaggcataggacccgtgtct / LE / cgtccatcagctcctggatcaTTTTTaggcataggacccgtgtct
LE / agcagggccacaaaggctTTTTTaggcataggacccgtgtct / LE / tgtgtcaaaggtttgcatctgagTTTTTaggcataggacccgtgtct
LE / caacagggcggatgaatctgTTTTTaggcataggacccgtgtct / LE / catgtggcagggtcttccaaTTTTTaggcataggacccgtgtct
LE / ttaagtagaggtttgctccaaacaTTTTTaggcataggacccgtgtct / LE / accctgtctttcatgatttgactcTTTTTaggcataggacccgtgtct
LE / cccacgctgatgttgcacaTTTTTaggcataggacccgtgtct / LE / gccgtcttctccgcgcaTTTTTaggcataggacccgtgtct
LE / gagccagatcctggtccagcTTTTTaggcataggacccgtgtct / LE / ttgacacatcggccaggtgTTTTTaggcataggacccgtgtct
BL / gtcatctggagcaggaagtcaa / BL / atcatctctttcttcatgccactc
BL / ggggcgagaggcctcc / BL / tccctcttcttcctcttgatcaa
BL / tgaaaagcagcacacgacgtc / BL / gcggtggagcctcaatcttt
BL / gggggcagatttcggct / BL / cagcccctgccctccag
BL / aggccttctttttgtttcggt / BL / gaaatccttgaagtgggagaaagt
BL / cttgcggaagaagcaaagtaag / BL / gaacactgcaggcagccg
BL / ggaccagaaagagcagcagca / BL / tggaagctcacagccactgtg
Npt1 (X77241) / BL / cagtgaggcctgcagaaactc
CE / ctgcctcgttctaagggaggtTTTTTctcttggaaagaaagt / BL / ggggttggtagttccagatgct
CE / tggcctgttgcatgagggaTTTTTctcttggaaagaaagt / BL / ttcccgtcgctcttggaag
CE / ggataatggcccagagtggaaTTTTTctcttggaaagaaagt / Oat5 (NM_144785)
LE / tgctggcctgttgacactgttTTTTTaggcataggacccgtgtct / CE / gggaagaatcccatagatgatccTTTTTctcttggaaagaaagt
LE / gcccatttgacccatatttcaTTTTTaggcataggacccgtgtct / CE / ttggtctcaggaagaagaagggTTTTTctcttggaaagaaagt
LE / aaccctgatagagtcatagaggtaagtTTTTTaggcataggacccgtgtct / CE / ttttcctcttattttctatatcttggatTTTTTctcttggaaagaaagt
LE / ggacaataaatggtcccattacaTTTTTaggcataggacccgtgtct / CE / ggccggttggaattctttataatTTTTTctcttggaaagaaagt
LE / tgctcatatacgggtggtccttTTTTTaggcataggacccgtgtct / LE / caacgaggccaccaaggaaTTTTTaggcataggacccgtgtct
LE / ggatatgatgtagtccttctcactgcTTTTTaggcataggacccgtgtct / LE / ggtgtcactttggcaaccacaTTTTTaggcataggacccgtgtct
LE / cagggattgtctgcctgagcTTTTTaggcataggacccgtgtct / LE / gaccacgatatgctgaatgatatatatTTTTTaggcataggacccgtgtct
LE / gagatttaagcatagctttgattggTTTTTaggcataggacccgtgtct / LE / ttatacatgacaaactagtataattttaatttaaTTTTTaggcataggacccgtgtct
LE / tgtatgtaaccaggagactgttcgTTTTTaggcataggacccgtgtct / LE / gattttattcaaatttctttattatagttattTTTTTaggcataggacccgtgtct
LE / gggaggctggacagcagtcTTTTTaggcataggacccgtgtct / BL / cgctcctccagcactgcca
LE / ccacagatgtaggcaagcagatagTTTTTaggcataggacccgtgtct / BL / cgtgaggatcataagtagaggagacag
LE / gtctgacatctgacctgctaggataTTTTTaggcataggacccgtgtct / BL / agggcaaggaagcagagtatgt
BL / cagatgaagccactcacaagca / BL / tgagtcaggcagaggctgattc
BL / gggccagcccagaagatca / BL / tctttctttgcctcttttgagc
BL / aacaataccaaagatgtagaagaccat / BL / tttttcatctttctccttgatcaga
BL / caggaaagactcagaacacaccc / BL / aaatcttgctaacatataggtataattctg
BL / ggggtcatcaaagaatagaaagaac / BL / ctcattatttatacagtgacttatatggaa
BL / accatatgaaagcaaagctattga / Ost alpha (NM_145932)
BL / cagtgctgataaatgtcggcg / CE / aggaatttgatcctctgtgctgtTTTTTctcttggaaagaaagt
BL / cattctctctaacattaacatgaagca / CE / ccttgttcagagctgggtgaagTTTTTctcttggaaagaaagt
PXR (AF031814) / CE / atccttcttttctctcaggcagaTTTTTctcttggaaagaaagt
CE / tggcagaagagggatgatctctTTTTTctcttggaaagaaagt / LE / gcccaggctcctgactcctcTTTTTaggcataggacccgtgtct
LE / tccaggcacttgcgcaaaTTTTTaggcataggacccgtgtct / LE / tttttatctcagagttcacccctgTTTTTaggcataggacccgtgtct
LE / acagcggcatcggacatgTTTTTaggcataggacccgtgtct / LE / ccacgcctgtttcattacctcTTTTTaggcataggacccgtgtct
Ost alpha (Cont’d) / Ost beta (Cont’d)
LE / gctaaacagtaagcggagcctTTTTTaggcataggacccgtgtct / LE / gatgtttctggccctggaccTTTTTaggcataggacccgtgtct
LE / cccgtattgaggctctcagagTTTTTaggcataggacccgtgtct / LE / gcagctttctctgcactgtggtTTTTTaggcataggacccgtgtct
LE / aggcagggacagccactgaTTTTTaggcataggacccgtgtct / LE / aataattccaaggagccgcaTTTTTaggcataggacccgtgtct
LE / cgctgaggacgaggagggtcTTTTTaggcataggacccgtgtct / LE / atgcttgtcatgaccaccaggTTTTTaggcataggacccgtgtct
BL / cccacagcttaggcgcg / LE / tctttcgatttctgtttgccagTTTTTaggcataggacccgtgtct
BL / ctcaagcctccagtgtcaagc / LE / catcaagatgcaggtcttctggtTTTTTaggcataggacccgtgtct
BL / ggggtggcgaggtttctgt / BL / cgggattcctgggacttca
BL / tatgagtcctgcctggctcc / BL / ggcacctcggcattggct
Ost beta (NM_178933) / BL / tcttctgcacgaaaataccaaag
CE / ccatgtttctgctcctgattgaTTTTTctcttggaaagaaagt / BL / actgccaggaccaggatgg
CE / catttcttccagcagttcctggTTTTTctcttggaaagaaagt / BL / gtttctttgtcttgtggctgct
CE / gatgctccttctcaggaggaacTTTTTctcttggaaagaaagt / BL / tcaatgtctctcttaggaagacctg
CE / gctgttgttctctttcattatggagtTTTTTctcttggaaagaaagt

Note:

aFunction refers to the type of bDNA oligonucleotide probe represented by each sequence.

bGenBank accession numbers for each transcript are given in cell under the gene name.

cCE, capture extender; LE, label extender; BL, blocker.

Fig. s1 mRNA expression of major hepatic transporters in male C57BL/6 and PXR-null mouse livers after administration of pregnenolone-16α-carbonitrile (PCN). Black bars represent WT mouse tissues; grey bars represent PCN-treated WT mouse tissues; latticed bars represent PXR-null mouse tissues; striated bars represent PCN-treated PXR-null mouse tissues. Four treatment groups were conducted: C57BL/6 mouse, C57BL/6 mouse-PCN, PXR-null mouse, and PXR-null mouse-PCN. Mice (n=5) were administered once daily for 4 days either corn oil ip as the control group, or with PCN (200mg/kg, ip in corn oil). After isolation of total RNA, pooled RNA samples or individual RNA samples of 5 mouse liver were analyzed by bDNA assay. Where individual RNA samples were quantified, data were expressed as mean±S.E.M. for five mice in each group. Asterisk (*) indicates statistically significant difference between treated and control mice (p<0.05). Dagger (†) indicates a statistically significant difference in constitutive transporter mRNA expression between C57BL/6 and PXR-null mouse liver.

Fig. s2 mRNA expression of major renal transporters in male C57BL/6 and PXR-null mouse kidneys after administration of pregnenolone-16α-carbonitrile (PCN). Black bars represent WT mouse tissues; grey bars represent PCN-treated WT mouse tissues; latticed bars represent PXR-null mouse tissues; striated bars represent PCN-treated PXR-null mouse tissues. Four treatment groups were conducted: C57BL/6 mouse, C57BL/6 mouse-PCN, PXR-null mouse, and PXR-null mouse-PCN. Mice (n=5) were administered once daily for 4 days either corn oil ip as the control group, or with PCN (200mg/kg, ip in corn oil). After isolation of total RNA, pooled RNA samples of 5 mouse kidneys were analyzed by the bDNA assay.

Fig. s3 mRNA expression of major duodenal transporters in male C57BL/6 and PXR-null mouse duodenums after administration of pregnenolone-16α-carbonitrile (PCN). Black bars represent WT mouse tissues; grey bars represent PCN-treated WT mouse tissues; latticed bars represent PXR-null mouse tissues; striated bars represent PCN-treated PXR-null mouse tissues. Four treatment groups were conducted: C57BL/6 mouse, C57BL/6 mouse-PCN, PXR-null mouse, and PXR-null mouse-PCN. Mice (n=5) were administered once daily for 4 days either corn oil ip as the control group, or with PCN (200mg/kg, ip in corn oil). After isolation of total RNA, pooled RNA sample or individual RNA samples of 5 mice were analyzed by bDNA assay. When individual RNA samples were quantified, data were expressed as mean±S.E.M. for five mice in each group. Asterisk (*) indicates statistically significant difference between treated and control mice (p<0.05). Dagger (†) indicates statistically significant difference in constitutive transporter mRNA expression between C57BL/6 and PXR-null mouse duodenum.

Fig. s4 mRNA expression of major jejunal transporters in male C57BL/6 and PXR-null mouse jejunums after administration of pregnenolone-16α-carbonitrile (PCN). Black bars represent WT mouse tissues; grey bars represent PCN-treated WT mouse tissues; latticed bars represent PXR-null mouse tissues; striated bars represent PCN-treated PXR-null mouse tissues. Four treatment groups were conducted: C57BL/6 mouse, C57BL/6 mouse-PCN, PXR-null mouse, and PXR-null mouse-PCN. Mice (n=5) were administered once daily for 4 days either corn oil ip as the control group, or with PCN (200mg/kg, ip in corn oil). After isolation of total RNA, pooled RNA samples or individual RNA samples of 5 mice were used for analysis by the bDNA assay. When individual RNA samples of each tissue were quantified, all data were expressed as mean±S.E.M. for five mice in each group. Asterisk (*) indicates statistically significant difference between treated and control mice (p<0.05). Dagger (†) indicates statistically significant difference in constitutive transporter mRNA expression of between C57BL/6 and PXR-null mouse jejunum.

Fig. s5 mRNA expression of major ileal transporters in male C57BL/6 and PXR-null mouse ileums after administration of pregnenolone-16α-carbonitrile (PCN). Black bars represent WT mouse tissues; grey bars represent PCN-treated WT mouse tissues; latticed bars represent PXR-null mouse tissues; striated bars represent PCN-treated PXR-null mouse tissues. Four treatment groups were conducted: C57BL/6 mouse, C57BL/6 mouse-PCN, PXR-null mouse, and PXR-null mouse-PCN. Mice (n=5) was administered once daily for 4 days either corn oil ip as the control or PCN (200mg/kg, ip in corn oil). After isolation of total RNA, pooled RNA sample or individual RNA samples of 5 mice were used for analysis by bDNA assay. When individual RNA sample of each tissue was used, all data are expressed as mean±S.E.M. for five mice in each group. Asterisk (*) indicates statistically significant difference between treated and control mice (p<0.05). Dagger (†) indicates statistically significant difference in constitutive transporter mRNA expression of between C57BL/6 and PXR-null mouse ileum.

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