Table 2.
qRT-PCR analysis to determine the mRNA decay half-life of representative late response genes. The cells were stimulated with growth factor ligands (EGF or HRG) for 4 h, and then ActD (mRNA synthesis inhibitor) or U0126 (MEK inhibitor). The mRNA decay rate (decay half-life) was derived as described in the Materials and methods. The data are the mean ± SD. NA, not applicable. The shortest duration of ligand/inhibitor combination in each cell type is shown in bold
| mRNA decay half-life (h) |
|||||||
|---|---|---|---|---|---|---|---|
| Gene | Cell type | Ligand | Ligand only | Ligand + ActD | Ligand + U0126 | mRNA stability | Number of ‘ATTTA’ motif |
| CDC42EP3 | WT | EGF | 8.58 ± 0.82 | 1.22 ± 0.2 | 1.62 ± 0.27 | ERK and transcription equally-dependent | 6 |
| HRG | 3.13 ± 0.05 | 1.26 ± 0.12 | 1.25 ± 0.16 | ||||
| 6KR | EGF | 4.55 ± 0.4 | 1.37 ± 0.07 | 1.38 ± 0.06 | |||
| HRG | 5.04 ± 0.13 | 1.72 ± 0.02 | 1.55 ± 0.04 | ||||
| DUSP6 | WT | EGF | NA | 2.31 ± 0.17 | 1.44 ± 0.07 | More ERK-dependent | 1 |
| HRG | 4.91 ± 0.91 | 1.02 ± 0.05 | 1.05 ± 0.21 | ||||
| 6KR | EGF | 2.81 ± 0.14 | 1.21 ± 0.01 | 1.07 ± 0.34 | |||
| HRG | 4.83 ± 0.06 | 1.73 ± 0.13 | 0.9 ± 0.13 | ||||
| EGFR | WT | EGF | > 20 | 4.97 ± 0.06 | 16.5 ± 6.09 | More transcription-dependent | 9 |
| HRG | 16.31 ± 0.35 | 6.99 ± 0.08 | 7.76 ± 0.1 | ||||
| 6KR | EGF | > 20 | 3.74 ± 0.1 | 16.98 ± 1.63 | |||
| HRG | NA | 4.24 ± 0.02 | > 20 | ||||
| FABP5 | WT | EGF | NA | > 20 | NA | Neither | 0 |
| HRG | NA | NA | NA | ||||
| 6KR | EGF | NA | NA | NA | |||
| HRG | 0.69 ± 0 | 0.69 ± 0 | 0.69 ± 0 | ||||
| FHL2 | WT | EGF | 4.37 ± 0.12 | 3.17 ± 0.01 | 3.5 ± 0.14 | More ERK-dependent | 6 |
| HRG | 4.3 ± 0.17 | 2.29 ± 0.08 | 2.19 ± 0.02 | ||||
| 6KR | EGF | 2.13 ± 0.1 | 1.77 ± 0.06 | 1.73 ± 0.03 | |||
| HRG | 2.56 ± 0 | 2.32 ± 0.1 | 1.96 ± 0.05 | ||||
| PDLIM7 | WT | EGF | > 20 | 6.54 ± 0.34 | 6.01 ± 0.69 | More ERK-dependent | 0 |
| HRG | > 20 | 8.23 ± 0.01 | 5.37 ± 0.1 | ||||
| 6KR | EGF | > 20 | 16.11 ± 0.46 | 5.01 ± 0.03 | |||
| HRG | NA | N.A | 15.16 ± 3.55 | ||||
| PRKCA | WT | EGF | NA | 10.21 ± 2.81 | NA | More transcription-dependent | 7 |
| HRG | NA | 4.06 ± 0.21 | NA | ||||
| 6KR | EGF | NA | NA | NA | |||
| HRG | NA | > 20 | NA | ||||
| SFN | WT | EGF | NA | 10.16 ± 0.29 | 6.31 ± 0.27 | More ERK-dependent | 0 |
| HRG | 6.18 ± 0.96 | 6.18 ± 0.91 | 4.63 ± 1.32 | ||||
| 6KR | EGF | 6.42 ± 0.5 | 7.55 ± 0.1 | 3.36 ± 0.12 | |||
| HRG | 17.37 ± 4.27 | > 20 | 7.72 ± 1.07 | ||||
| SPHK1 | WT | EGF | > 20 | 3.9 ± 0.51 | 4.77 ± 0.14 | More transcription-dependent | 0 |
| HRG | > 20 | 2.76 ± 0.88 | 7.18 ± 2.86 | ||||
| 6KR | EGF | 11.63 ± 15.47 | 3.47 ± 3.93 | 4.48 ± 5.35 | |||
| HRG | 6.02 ± 7.54 | 3.58 ± 4.09 | 6.87 ± 8.74 | ||||
| ZYX | WT | EGF | 7.34 ± 1.24 | 4.47 ± 0.41 | 5.55 ± 1.4 | More ERK-dependent | 0 |
| HRG | 7.87 ± 2.06 | 8.29 ± 3.87 | 6.6 ± 2.54 | ||||
| 6KR | EGF | 4.13 ± 0.69 | 4.7 ± 0.78 | 3.09 ± 0.66 | |||
| HRG | 2.57 ± 2.65 | 3.55 ± 4.04 | 2.45 ± 2.49 | ||||