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Table 7 Summary of mRNA levels in GFP shRNA control transfectants, URE3-BP shRNA transfectants, and nontransfected HM1:IMSS trophozoites

From: Short hairpin RNA-mediated knockdown of protein expression in Entamoeba histolytica

shRNA transfectant or control sample URE3-BP 5' oligo pair P-value URE3-BP middle oligo pair P-value URE3-BP 3' oligo pair P-value
GFP 100.0 ± 2.9 -- 100 ± 2.8 -- 100 ± 4.3 --
HM1:IMSS 106.4 ± 5.8 0.2928 108.9 ± 5.6 0.1008 102.8 ± 5.0 0.5792
URE3-BP (350–378) 67.0 ± 2.5 <0.0001 67.4 ± 2.0 <0.0001 72.2 ± 2.8 <0.0001
URE3-BP (580–608) 12.4 ± 0.8 <0.0001 13.5 ± 3.3 <0.0001 12.5 ± 3.8 <0.0001
  1. The average URE3-BP transcript level as measured by qRT-PCR and normalized to actin was defined as being 100% in the GFP shRNA control transfectants. HM1:IMSS nontransfected amebae were also included. Three different oligo pairs amplifying the 5', middle, and 3' sections of URE3-BP were used (sequences and locations are shown in Table 3). Student's t test was used for statistical analysis. Three biological replicates were each assayed in quadruplicate with each oligo pair, with the exception of the HM1:IMSS samples, which had one biological replicate. Values are expressed as the percentage of URE3-BP mRNA of the GFP control shRNA transfectant level ± SE, with the P-value following each.