Table 1.
Quantitative polymerase chain reactions (qPCRs) were performed in the C1000 Thermal Cycler coupled to a CFX96 Real Time System (Bio-Rad, Australia).
| Gene | Fwd/Rev | Sequence 5′ to 3′ | Melt ∘C |
|---|---|---|---|
| TH | Fwd | CGG GCT TCT CGG ACC AGG TGT A | 73.2 |
| Rev | CTC CTC GGC GGT GTA CTC CAC A | 72.6 | |
| AADC | Fwd | CTC GGA CCA AAG TGA TCC AT | 63.9 |
| Rev | GTC TCT CTC CAG GGC TTC CT | 63.8 | |
| NURR1 | Fwd | TTC TCC TTT AAG CAA TCG CCC | 66.1 |
| Rev | AAG CCT TTG CAG CCC TCA CAG | 69.8 | |
| PITX3 | Fwd | GCC AAC CTT AGT CCG TG | 59.2 |
| Rev | GCA AGC CAG TCA AAA TG | 57.8 | |
| DAT | Fwd | AGC AGA ACG GAG TGC AGC T | 65.4 |
| Rev | GTA TGC TCT GAT GCC GTC T | 60.5 | |
| FOXG1 | Fwd | TGG CCC ATG TCG CCC TTC CT | 75.1 |
| Rev | GCC GAC GTG GTG CCG TTG TA | 73.5 | |
| NKX2.1 | Fwd | AGG GCG GGG CAC AGA TTG GA | 75.1 |
| Rev | GCT GGC AGA GTG TGC CCA GA | 71.4 | |
| PAX5 | Fwd | CCC CAT TGT GAC AGG CCG TGA C | 74.8 |
| Rev | TCA GCG TCG GTG CTG AGT AGC T | 70.5 | |
| EN1 | Fwd | TCT CGC TGT CTC TCC CTC TC | 63.9 |
| Rev | CGT GGC TTA CTC CCC ATT TA | 63.6 | |
| WNT1 | Fwd | GAG CCA CGA GTT TGG ATG TT | 64.0 |
| Rev | TGC AGG GAG AAA GGA GAG AA | 64.0 | |
| GIRK2 | Fwd | GCT ACC GGG TCA TCA CAG AT | 63.9 |
| Rev | ACT GCA TGG GTG GAA AAG AC | 63.9 | |
| LMX1A | Fwd | CGC ATC GTT TCT TCT CCT CT | 63.4 |
| Rev | CAG ACA GAC TTG GGG CTC AC | 65.0 | |
| FOXA2 | Fwd | GGG GTA GTG CAT CAC CTG TT | 63.8 |
| Rev | CCG TTC TCC ATC AAC AAC CT | 63.9 | |
| SHH | Fwd | CCA ATT ACA ACC CCG ACA TC | 63.9 |
| Rev | AGT TTC ACT CCT GGC CAC TG | 64.3 | |
| CORIN | Fwd | CAT ATC TCC ATC GCC TCA GTT G | 65.6 |
| Rev | GGC AGG AGT CCA TGA CTG T | 62.9 | |
| HOXA2 | Fwd | CGT CGC TCG CTG AGT GCC TG | 74.1 |
| Rev | TGT CGA GTG TGA AAG CGT CGA GG | 72.7 | |
| ACTB | Fwd | CCT TGC ACA TGC CGG AG | 66.9 |
| Rev | GCA CAG AGC CTC GCC TT | 64.6 | |
| GAPDH | Fwd | TTG AGG TCA ATG AAG GGG TC | 63.9 |
| Rev | GAA GGT GAA GGT CGG AGT CA | 64.6 | |
| OCT4 | Fwd | TCT CCA GGT TGC CTC TCA CT | 64.1 |
| Rev | GTG GAG GAA GCT GAC AAC AA | 62.9 | |
| MAP2 | Fwd | CCG TGT GGA CCA TGG GGC TG | 75.3 |
| Rev | GTC GTC GGG GTG ATG CCA CG | 75.4 | |
| SYP | Fwd | ACC TCG GGA CTC AAC ACC TCG G | 72.6 |
| Rev | GAA CCA CAG GTT GCC GAC CCA G | 73.8 |
Reaction conditions included cDNA synthesis (10 min at 50∘C), reverse transcriptase inactivation (5 min at 95∘C), PCR cycling (10 s at 95∘C then 30 s at 55–60∘C; up to 43 cycles), and melt curve analysis (1 min at 95∘C, 1 min at 55∘C, and 10 s each at steps of 0.5∘C between and 55–95∘C). Primers used in gene expression assay.