Table 2B.
Primers and probes for quantitative real-time PCR.
| PCR assay primer (annealing temperature) | Oligonucleotide sequence (5′→3′) | Amplicon length (Bp) | GenBank No. |
|---|---|---|---|
| At1a (55°C) | |||
| Agtr1 S | GCAAAGCTGTCTTACATTAATAGAT | 121 | NM_177322 |
| Agtr1 A | AATCAAAAGGAGACCGCT | ||
| Agtr1 FL | CCACAAATCCATCCAgCTCCT-FL | ||
| Agtr1 LC 640 | Red640-ACTTGTCCTTGGGGCAGCCA-Ph | ||
| At1b (55°C) | |||
| Agtr1b F | GTGTGTTAAGATTTGCTAGGC | 208 | NM_175086 |
| Agtr1b A | TgAAATAAAACTGGTCCAACTAT | ||
| Agtr1b FL | CATGCATTACCTCAGTCATAAAGTCAA–FL | ||
| Agtr1b LC 640 | Red640-CTGCTGTGATTCTCTCCCAGGT–PH | ||
| At2 (55°C) | |||
| Agtr2 S | GGGGAGTAGTTTGAATCTGC | 154 | NM_007429 |
| Agtr2 A | GGTAACACAGCTTCAGGTATTATATT | ||
| Agtr2 FL | TCGTGTTAAGAATGAGTTCTGTGGACC-FL | ||
| Agtr2 LC 640 | Red640-GGGCTTGCTTTAAATCACCTTCACAG-PH | ||
| eNos (55°C) | |||
| eNos F | TCCGATTCAACAGTGTCTCC | 306 | NM_008713 |
| eNos R | CCACACAGAAGGTTTCACAG | ||
| eNos FL | TCCGATT CAACA GTGTCT CCTGC-FL | ||
| eNos Cy5 | Cy5-TCAGACCCACTGGTATCCTCTTGG-PH | ||
| Arg1 (58°C) | |||
| Arg1 F | CTCCAAGCCAAAGTCCTTAGAG | 185 | NM_007482 |
| Arg1 R | AGGAGCTGTCATTAGGGACATC | ||
| Ym1 (58°C) | |||
| Ym1 F | CAGGTCTGGCAATTCTTCTGAA | 197 | M94584 |
| Ym1 R | GTCTTGCTCATGTGTGTAAGTGA |
Mouse specific primers and probes and optimized temperature conditions for quantitative real-time polymerase chain reaction (qPCR): angiotensin II receptors type 1A (At1a), 1B (At1b) and 2 (At2), arginase 1 (Arg1), chitinase-like protein 3 (Ym1), endothelial nitric oxide synthase (eNos); F: forward, R: reverse. Kits (manufacturer): Ppia: Absolute Fast qPCR Mix AB-4325 (Abgene); Agtr1 (At1a), Agtr1b (At1b), Agtr2 (At2): Maxima Probe qPCR Master Mix K0262 (Fermentas); Il1b, Il6, eNos, iNos: Light Cycler 480 Probes Master 04887301001 (Roche); Tnfa, Arg1 and Ym1: Absolute Blue Qpcr Sybrgreenmix AB-4166 (Thermo Scientific).