FIG. 10.

Outcome and mechanism of enhanced ccn2 gene expression by npm gene silencing. (A) Cellular CCN2 levels in CEFs as evaluated by Western blotting after the introduction of the siRNAs targeting npm. The same set of siRNAs was employed as in Fig. 9. The same amount of total proteins (10 μg) was loaded for each sample. (B) No significant effect of npm gene silencing on the degradation of gapdh mRNA in vivo. Nascent mRNA synthesis was arrested by actinomycin D treatment, and the fate of the mRNA in CEFs with a control RNA duplex (solid line) or si546 (dotted line) was pursued by real-time RT-PCR analysis. Relative values were computed against those at time zero. (C) Effect of npm gene silencing on the degradation of ccn2 mRNA in CEFs transfected with a control RNA duplex (solid line) or si546 (dotted line). Standardized values of ccn2 mRNA levels (ccn2/gapdh) were further normalized against those at time zero and presented.