Screening and validation of siRNAs in mouse primary hepatocytes.
Screening of siRNA in mouse primary hepatocytes 24 hours after transfection with 0.3, 3 and 30nM of three Hnf4a-specific (#1–3, panel A), three Cebpa -specific siRNAs (#1–3, panel B) or a control siRNA (siNEG, panel A and B, open bars). Hnf4a and Cebpa transcript levels were determined by quantitative real-time PCR. β-actin was used as internal control for quantification and normalization. The ΔCt values of the individual samples were related to the mean ΔCt of the reference group (siNEG). On the x-axis siRNA concentrations are indicated. Data are expressed as mean with error bars representing the difference between 2 POWER of upper and lower range of the mean ΔΔCt (difference 2∧ΔΔCt+SEM and 2∧ΔΔCt−SEM). Individual experiments were performed in triplicate. Data were statistically analyzed using the Student's t-test. P-values <0.05 were regarded as statistically significant. *P<0.05, **P<0.01, ***P<0.001.