Gene expression measured by qRT-PCR
| Hsp70_5 | ||||
| CTRL | HS | |||
| Endogenous | Transgene | Endogenous | Transgene | |
| aRelative | 1 | 0.63 ± 0.11 | 140 ± 23 | 200 ± 56 |
| bNormalized | 1 | 0.14 ± 0.02 | 140 ± 23 | 45 ± 12 |
| Hsp70_5 | ||||
| CTRL | HS | |||
| Endogenous | Transgene | Endogenous | Transgene | |
| aRelative | 1 | 0.63 ± 0.11 | 140 ± 23 | 200 ± 56 |
| bNormalized | 1 | 0.14 ± 0.02 | 140 ± 23 | 45 ± 12 |
| MT1_1_4 | DHFR D10 | ||||
| CTRL | Zn | Log phase | G1 block | ||
| 2 h | 16 h | (CTRL) | |||
| cRelative | 1 | 3.9 ± 0.14 | 51 ± 8.3 | 1 | 0.31 ± 0.046 |
| MT1_1_4 | DHFR D10 | ||||
| CTRL | Zn | Log phase | G1 block | ||
| 2 h | 16 h | (CTRL) | |||
| cRelative | 1 | 3.9 ± 0.14 | 51 ± 8.3 | 1 | 0.31 ± 0.046 |
Expression levels relative to endogenous HSPA1A expression level at control temperature—human specific primers were used for BAC transgene, a 100% rat/mouse conserved sequence (specific for the CHO endogenous HSPA1A copy) was used for the endogenous gene.
Expression levels normalized for copy number estimated by mean of qPCR data (Table I).
Expression levels relative to control.
qRT-PCR was performed on four independent experiments for the Hsp70_5 cell line and three independent experiments for the DHFR D10 and MT1_1_4 cell lines. The comparative 2−ΔΔCt method was used for relative expression level measurements. The β-actin gene was used as the endogenous control to normalize RNA loading. Numbers shown are the mean ± SE. PCR primers are listed in Table S2.