Table 1. Primer pairs and TaqMan probes used in qPCR for Comparative Gene Expression in Aggregates and Control Monolayer HepG2.
Primer and probes for qPCR were designed using Roche Universal Probe Library Assay Design Center (https://www.roche-applied-science.com). %GC content was maintained between 45% and 55%. Designed primers were obtained from Eurofins MWG Operon (http://www.eurofinsdna.com), and TaqMan probes were purchased from Roche.
| Gene | Gene Symbol | Primer Sequence | Roche Library Probe No. | Accession No. |
|---|---|---|---|---|
| Solute carrier organic anion transporter family, member 1B1 (SLCO1B1) | OATP1B1 | gttccatcattcatatagaacggaga (f) | 35 | NM_006446.4 |
| tcaaagcttccgtcaataaaacc (r) | ||||
| Solute carrier organic anion transporter family, member 1B3 (SLCO1B3) | OATP1B3 | catgtaatttggacatgcaagaca | 74 | NM_019844.2 |
| aacatcttaatgaatcaatgcaatgttag | ||||
| Solute carrier organic anion transporter family, member 1A2 (SLCO1A2) | OATP1A2 | ctactttctggccttatgccatc | 42 | NM_134431.3 |
| agatggattccaggtttgaggat | ||||
| Solute carrier organic anion transporter family, member 2B1 (SLCO2B1) | OATP2B1 | agaggctggacttgaacaaacc | 36 | NM_001145212.2 |
| tgtttgtctccctgggtcactt | ||||
| Cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) | CYP1A1 | cctgggaacatcacattcctct | 4 | NM_000499.3 |
| agaaggcagccctgtttgttc | ||||
| Glutathione peroxidase 1 | GPX | tctgttgctcgtagctgctgaa | 55 | ENST00000419349.1 |
| ctatgagtcaccgggattttgc | ||||
| GSTA1-001 glutathione S-transferase alpha 1 | GST | tgggagagaactattgagaggaaca | 85 | ENST00000334575.5 |
| tgttaaacgctgtcaccgtcc | ||||
| Human multidrug resistance protein (MDR1) gene enhancer | MDR1 | tttcctggttagtcaatgggaga | 6 | M57451.1 |
| ggtgctcttggtttgctcaca | ||||
| Human P-glycoprotein gene (MDR2), unknown exon, clone pHDR2.6 | MDR2 | tgaagggcctcaacctgaag | 81 | M28477.1 |
| tcttcccacagccactacttcc | ||||
| Multidrug resistance-associated protein (MRP)-likeprotein-2 (MLP-2) | MLP-2 | cctggctgtgctctacacct | 5 | AB010887.1 |
| tgattccagccgcttcagtt | ||||
| Glyceraldehyde-3-phosphate dehydrogenase (GADPH) | GADPH | ctctgctcctcctgttcgaca | 60 | AF261085.1 |
| aatccgttgactccgaccttc | ||||
| Topoisomerase (DNA) III beta (TOP3B) | TOP3B | tcttgctgtgggggttcag | 39 | NM_003935.3 |
| tgggagggttcgaagaagaa | ||||
| RNA polymerase II, TATA box binding protein (TBP)-associated factor | TBP | ggtaaccactgttccgaagcc | 5 | NM_005640.1 |
| actgtgacgacactggttgg | ||||
| Elongation factor 2 (EF-2) | EF-2 | aaaggaacacgcgggtagatg | 4 | M19997.1 |
| tcatggtgacaccgcacag | ||||
| Elongation factor-1 alpha (EF-1) | EF-1 | ggaccattgagaagttcgagaag | 60 | L10340.1 |
| agcacccaggcatacttgaag |