TABLE 1.

Determination of gene copy number

DNA concentrations were measured by absorbance at 260 nm. Standard curves of C_T versus log DNA concentration were obtained for endogenous Eif3e and GAPDH genes by performing qPCR on DNA from 3T3-Ø and 3T3-3e5 cells with primer sets 8 and 10, respectively (supplemental Table S1). Sequences from Eif3e and GAPDH genes were amplified in quintuplicate from 16 ng of DNA for each cell line. The C_T values were used to calculate the concentrations of these sequences from the appropriate standard curve. The concentration of Eif3e sequences was then normalized by the concentration of GAPDH sequences for a given cell line. Finally, the copy number of Eif3e in 3T3-3e5 DNA was calculated relative to the copy number in 3T3-Ø DNA.

Cell Line	Eif3e	GAPDH	Eif3e normalized to GAPDH	Eif3e copy number normalized to 3T3-Ø
3T3-Ø	17.45	14.33
	16.51	14.41
	18.64	13.82
	21.51	13.97
	21.99	19.28
Average	19.22	15.16	1.27	1
3T3-3e5	39.01	16.41
	27.26	12.78
	30.95	15.34
	24.55	14.79
	23.24	10.55
Average	29.00	13.97	2.08	1.64 ± 0.21