Table 1.
shRNA construct | Log10 reduction valuea and CPE scoreb | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
pENTR/U6-shRNA (μg) | ||||||||||||
0.25 | 0.5 | 0.75 | 1 | |||||||||
qRT-PCR | TCID50 | CPE | qRT-PCR | TCID50 | CPE | qRT-PCR | TCID50 | CPE | qRT-PCR | TCID50 | CPE | |
Ni1 | 1.35a | 1.17a | 3.0 | 2.14b | 1.67b | 2.0 | 2.23b | 3.28c | 1.3 | 2.65c | 3.28c | 1.0 |
Ni2 | 0.71a | 1.22a | 3.0 | 1.48b | 2.22b | 2.3 | 1.92c | 3.00b | 1.8 | 2.53d | 3.00b | 1.0 |
Ni3 | 1.12a | 1.28a | 3.0 | 1.63b | 2.44b | 2.3 | 2.03bc | 2.78b | 1.3 | 2.29c | 2.78b | 1.0 |
Li1 | ns | ns | 4.0 | 0.29a | 0.61a | 3.0 | 0.27ab | 0.50a | 3.0 | 0.41b | 0.78a | 2.0 |
Li2 | 0.26a | 0.50a | 3.0 | 0.36ab | 0.56a | 3.0 | 0.39b | 0.50a | 3.0 | 0.31ab | 0.56a | 2.3 |
Li3 | ns | ns | 4.0 | 0.12a | ns | 3.0 | 0.17a | ns | 3.0 | 0.27b | 0.72a | 2.0 |
SCR | ns | ns | 4.0 | ns | ns | 4.0 | ns | ns | 4.0 | ns | ns | 4.0 |
Ni shRNAs associationc | Ni1 (1.2 μg) | Ni2 (1.2 μg) | Ni3 (1.2 μg) | Ni1 + Ni2 + Ni3 (0.4 μg each) | ||||||||
2.54b | 3.1a | 1.0 | 1.90a | 3.1a | 1.0 | 1.92a | 3.1a | 1.0 | 1.87a | 3.1a | 1.0 |
aThe log10 reduction was calculated by subtracting the log10 means of the CDV infectivity in the presence of transfected shRNA constructs from the log10 means of the CDV infectivity in the absence of shRNAs
bCPE scores were defined as (1) CPE < 25%, (2) CPE between 25 and 50%, (3) CPE between 51 and 75% and (4) CPE > 75%. CPE values correspond to the CPE score means for replicates
cAssociation of pENTR/U6-shRNAs against CDV nucleoprotein
Values followed by the same lowercase letters in horizontal lines do not differ within each method; p < 0.01 (Tukey’s test)
Ni plasmid shRNA for CDV nucleoprotein mRNA, Li plasmid shRNA for CDV large protein mRNA, SCR plasmid scramble shRNA, ns (not statistically significant): no significant difference between shRNA-treated and untreated CDV infected cells