TABLE V.
Real-Time PCR Detection of IL-8 mRNA in HUVEC Infected With Lassa and Mopeia Virusesa
| Threshold Cycle (CT):
|
40/20 hours:
|
|||
|---|---|---|---|---|
| Infection | IL-8 mRNA | GAPDH mRNA | ΔCT IL-8b | ΔIL-8 |
| HUVEC-mock | 26.55 ± 0.20 | 32.42 ± 0.24 | ||
| HUVEC-Lassa-20 | 26.78 ± 0.26 | 31.65 ± 0.36 | ||
| HUVEC-Lassa-40 | 29.40 ± 0.03 | 32.95 ± 0.16 | (−)1.32 | (−)2.5× |
| HUVEC-Mopeia-20 | 27.43 ± 0.08 | 30.64 ± 0.29 | ||
| HUVEC-Mopeia-40 | 29.50 ± 0.49 | 32.84 ± 0.30 | 0.13 | 0.91× |
a
cDNAs were made from RNA extracted from mock or Lassa-, Mopeia-infected cells at 20 and 40 hours p. i. and amplified with IL-8 and GAPDH primers using the GeneAmp 5700 System with SYBR Green I chemistry. Amplification plots were analyzed with 5700 SDS software and expressed as CT values. The CT is a PCR cycle at which a statistically significant increase in fluorescence can be detected above the baseline. All PCR reactions had the identical dissociation curves (not shown).
b
ΔCT IL-8 values were normalized and subtracted to HUVEC-mock samples (see text for details).