TABLE 2.
Effect of E4F262 expression of E1A transformation of primary REFs
| Expression construct | Coexpressed oncogene(s) | Fold change in no. of REF foci (SD)a |
|---|---|---|
| pCMV5 | E1A(13S) + T24 ras | 1.00 |
| E4F262b | E1A(13S) + T24 ras | 0.05 (0.01) |
| E4F262 | E1A(12S) + T24 ras | 0.35 (0.05) |
| E4F-N1 | E1A(12S) + T24 ras | 0.37 (0.03) |
| E4F-N2 | E1A(12S) + T24 ras | 0.90 (0.04) |
| E4F-N5 | E1A(12S) + T24 ras | 0.86 (0.01) |
| E4F-C3 | E1A(12S) + T24 ras | 0.89 (0.04) |
| E4F-N1(fs) | E1A(12S) + T24 ras | 1.18 (0.07) |
| NF-IL6 | E1A(12S) + T24 ras | 1.13 (0.02) |
Calculated as number of transformed foci from transfection containing pCMV construct expressing E4F262, E4F262 mutant, NF-IL6, or ATF-2/number of foci from parallel pCMV5 control transfections. Values were derived from two independent experiments, each performed in duplicate or triplicate. pCMV5 control transfections containing E1A and T24 ras expression constructs typically produced 100 to 200 transformed foci per plate. pCMV5 or pCMVs-E4F262 transfections that did not contain E1A and T24 ras expression constructs produced zero to four foci per plate. A focus diameter of ≥2 mm was used as the criterion for transformation.
Each transfection of 105 passage 3 REFs contained 1.25 μg of pCMV5 or the indicated pCMV-transcription factor construct, 1.25 μg of pCMV-E1A(13S) or pCMV-E1A(12S), 1.25 μg of pSP72-ras (T24 ras), and sonciated salmon sperm DNA, for a total of 20.0 μg of DNA.