Semiquantitative RT-PCR measurement of gene expression in rat tissues including a correction for varying cell size and number

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.

ISSN: 1743-7075