Table 4.
Comparison of results obtained using a delta-delta (after correction using constitutive genes) method, quantitative RT-PCR approach (individual cDNA standard curves) and our postulated method (common cDNA bp-based standard curve) on liver and WAT from animals with restricted access to food and their controls.
ORGAN and gene | Group | Delta-delta | Individual RT-PCR | Present method |
LIVER | ||||
Carbohydrate-responsive element-binding protein | Control | 100 ± 4 | 100 ± 5 | 100 ± 5 |
Restricted | 76 ± 4 | 79 ± 4 | 79 ± 4 | |
Acetyl-CoA carboxylase 1 | Control | 100 ± 8 | 100 ± 3 | 100 ± 3 |
Restricted | 50 ± 12 | 47 ± 21 | 46 ± 1 | |
Insulin receptor substrate 1 | Control | 100 ± 8 | 100 ± 9 | 100 ± 8 |
Restricted | 116 ± 10 | 115 ± 10 | 115 ± 10 | |
ADIPOSE TISSUE | ||||
Carbohydrate-responsive element-binding protein | Control | 100 ± 12 | 100 ± 10 | 100 ± 9 |
Restricted | 24 ± 4 | 27 ± 5 | 27 ± 5 | |
Type II iodothyronine deiodinase | Control | 100 ± 6 | 100 ± 6 | 100 ± 5 |
Restricted | 45 ± 3 | 46 ± 2 | 46 ± 2 | |
Acetyl-CoA carboxylase 1 | Control | 100 ± 8 | 100 ± 13 | 100 ± 11 |
Restricted | 55 ± 14 | 53 ± 14 | 53 ± 11 | |
Insulin receptor substrate 1 | Control | 100 ± 13 | 100 ± 12 | 100 ± 11 |
Restricted | 28 ± 5 | 29 ± 5 | 28 ± 4 |
The data are the mean ± sem of 7 different animals and are expressed in all the cases as the percentage of the mean controls' values for easier comparison between the methods using homologous units. The base data used for comparisons in the case of the postulated method (and the individual quantitative RT-PCR method) was fmol/unit of total tissue RNA weight.