Table 1.
qRT-PCR data demonstrating specific EhMSP-1 silencing in G3 strain E. histolytica trophozoitesa
| Time after G418 removal | Gene | Sample | CT (mean ± SD) | ΔCT | ΔΔCT | Fold difference |
|---|---|---|---|---|---|---|
| 1 mo | EhMSP-1 | G3 control | 19.4 ± 0.1 | 0.5 | ||
| G3 EhMSP-1 (−) | 31.3 ± 0.3 | 12.6 | 12.1 | 4,271.2 | ||
| EhMSP-2 | G3 control | 32.1 ± 1.0 | 13.2 | |||
| G3 EhMSP-1 (−) | 31.8 ± 0.7 | 13.0 | −0.2 | 0.9 | ||
| L10 | G3 control | 18.9 ± 0.3 | ||||
| G3 EhMSP-1 (−) | 18.8 ± 0.6 | |||||
| 1 yr | EhMSP-1 | G3 control | 19.7 ± 0.1 | 1.3 | ||
| G3 EhMSP-1 (−) | 31.3 ± 0.5 | 12.7 | 11.4 | 2,814.4 | ||
| EhMSP-2 | G3 control | 31.3 ± 0.6 | 12.9 | |||
| G3 EhMSP-1 (−) | 31.3 ± 0.5 | 12.8 | −0.1 | 0.9 | ||
| L10 | G3 control | 18.4 ± 0.2 | ||||
| G3 EhMSP-1 (−) | 18.6 ± 0.3 |
a
Trophozoite mRNA was isolated from the EhMSP-1-silenced (−) and empty vector control G3 strain trophozoites 1 month and 1 year after removal of G418 selection. cDNA was synthesized using oligo(dT) primers, and gene transcripts for EhMSP-1, EhMSP-2, and L10 (ribosomal protein) were quantified by qRT-PCR using SYBR green. Shown are the mean and SD for threshold cycle numbers (CT). The fold change in gene expression for EhMSP-1 and EhMSP-2 was calculated using the comparative CT method and L10 as the control gene (34).