Effect of hGATA-1, hGATA-2, and hGATA-3 Transcription Factors on Induction of the GH Reporter in Hep3B Cells
| No. . | Plasmid DNA . | O2 . | hGH (ng)/CAT (U) . | |||
|---|---|---|---|---|---|---|
| . | . | (%) . | Exp 1 . | Exp 2 . | Exp 3 . | Exp 4 . |
| 1 | XGH | 21 | 0.483 | 4.428 | 5.871 | 5.238 |
| 2 | XGH | 1 | 0.641 | 3.987 | 4.568 | 2.250 |
| 3 | XGH + hGATA-1 | 21 | 0.284 | 6.245 | 7.597 | 5.375 |
| 4 | XGH + hGATA-1 | 1 | 1.697 | 5.859 | 10.116 | 8.805 |
| 5 | XGH + hGATA-2 | 21 | 0.620 | 3.844 | 2.881 | 3.837 |
| 6 | XGH + hGATA-2 | 1 | 0.760 | 2.625 | 9.298 | 1.639 |
| 7 | XGH + hGATA-3 | 21 | 0.896 | 4.039 | 7.621 | 1.089 |
| 8 | XGH + hGATA-3 | 1 | 1.301 | 1.231 | 3.856 | 1.673 |
| 9 | 5AXGH | 21 | 0.417 | 1.190 | 12.994 | 0.938 |
| 10 | 5AXGH | 1 | 0.278 | 0.798 | 3.001 | 0.763 |
| 11 | 5AXGH + hGATA-1 | 21 | 0.190 | 3.201 | 5.274 | 1.461 |
| 12 | 5AXGH + hGATA-1 | 1 | 0.108 | 1.357 | 4.857 | 2.804 |
| 13 | 5AXGH + hGATA-2 | 21 | 0.189 | 4.164 | 8.707 | 2.775 |
| 14 | 5AXGH + hGATA-2 | 1 | 0.022 | 0.572 | 9.465 | 0.952 |
| 15 | 5AXGH + hGATA-3 | 21 | 0.287 | 3.495 | 7.764 | 1.287 |
| 16 | 5AXGH + hGATA-3 | 1 | 0.026 | 0.383 | 7.207 | 0.553 |
| 17 | 5AXGH3Ac | 21 | 1.117 | 0.591 | 1.999 | 1.037 |
| 18 | 5AXGH3Ac | 1 | 1.324 | 0.723 | 3.904 | 1.714 |
| 19 | 5AXGH3Ac + hGATA-1 | 21 | 1.605 | 4.608 | 1.495 | 2.764 |
| 20 | 5AXGH3Ac + hGATA-1 | 1 | 0.480 | 1.065 | 1.272 | 3.822 |
| 21 | 5AXGH3Ac + hGATA-2 | 21 | 0.440 | 0.780 | 4.382 | 10.816 |
| 22 | 5AXGH3Ac + hGATA-2 | 1 | 0.193 | 0.335 | 2.366 | 4.608 |
| 23 | 5AXGH3Ac + hGATA-3 | 21 | 0.414 | 0.731 | 3.284 | 6.347 |
| 24 | 5AXGH3Ac + hGATA-3 | 1 | 0.098 | 0.197 | 1.189 | 2.978 |
| No. . | Plasmid DNA . | O2 . | hGH (ng)/CAT (U) . | |||
|---|---|---|---|---|---|---|
| . | . | (%) . | Exp 1 . | Exp 2 . | Exp 3 . | Exp 4 . |
| 1 | XGH | 21 | 0.483 | 4.428 | 5.871 | 5.238 |
| 2 | XGH | 1 | 0.641 | 3.987 | 4.568 | 2.250 |
| 3 | XGH + hGATA-1 | 21 | 0.284 | 6.245 | 7.597 | 5.375 |
| 4 | XGH + hGATA-1 | 1 | 1.697 | 5.859 | 10.116 | 8.805 |
| 5 | XGH + hGATA-2 | 21 | 0.620 | 3.844 | 2.881 | 3.837 |
| 6 | XGH + hGATA-2 | 1 | 0.760 | 2.625 | 9.298 | 1.639 |
| 7 | XGH + hGATA-3 | 21 | 0.896 | 4.039 | 7.621 | 1.089 |
| 8 | XGH + hGATA-3 | 1 | 1.301 | 1.231 | 3.856 | 1.673 |
| 9 | 5AXGH | 21 | 0.417 | 1.190 | 12.994 | 0.938 |
| 10 | 5AXGH | 1 | 0.278 | 0.798 | 3.001 | 0.763 |
| 11 | 5AXGH + hGATA-1 | 21 | 0.190 | 3.201 | 5.274 | 1.461 |
| 12 | 5AXGH + hGATA-1 | 1 | 0.108 | 1.357 | 4.857 | 2.804 |
| 13 | 5AXGH + hGATA-2 | 21 | 0.189 | 4.164 | 8.707 | 2.775 |
| 14 | 5AXGH + hGATA-2 | 1 | 0.022 | 0.572 | 9.465 | 0.952 |
| 15 | 5AXGH + hGATA-3 | 21 | 0.287 | 3.495 | 7.764 | 1.287 |
| 16 | 5AXGH + hGATA-3 | 1 | 0.026 | 0.383 | 7.207 | 0.553 |
| 17 | 5AXGH3Ac | 21 | 1.117 | 0.591 | 1.999 | 1.037 |
| 18 | 5AXGH3Ac | 1 | 1.324 | 0.723 | 3.904 | 1.714 |
| 19 | 5AXGH3Ac + hGATA-1 | 21 | 1.605 | 4.608 | 1.495 | 2.764 |
| 20 | 5AXGH3Ac + hGATA-1 | 1 | 0.480 | 1.065 | 1.272 | 3.822 |
| 21 | 5AXGH3Ac + hGATA-2 | 21 | 0.440 | 0.780 | 4.382 | 10.816 |
| 22 | 5AXGH3Ac + hGATA-2 | 1 | 0.193 | 0.335 | 2.366 | 4.608 |
| 23 | 5AXGH3Ac + hGATA-3 | 21 | 0.414 | 0.731 | 3.284 | 6.347 |
| 24 | 5AXGH3Ac + hGATA-3 | 1 | 0.098 | 0.197 | 1.189 | 2.978 |
Hep3B cells were transfected with each of the Epo-GH constructs and hGATA transcription factors hGATA-1, hGATA-2, and hGATA-3, as well as RSVCAT as an internal standard. At the end of a 4-day incubation in either 1% O2 (hypoxia) or 21% O2 (control), GH was measured in the cell media, and the cell pellet was assayed for CAT. To correct for variations in transfection efficiency, these results were normalized for expression of CAT. Four separate experiments were performed (n = 4).