FIGURE 7.

Circular RNAs are not associated with ribosomes and are susceptible to siRNA knockdown. Jurkat cell lysates were separated by sucrose gradient centrifugation. (A) Agarose gel to verify separation of 40S, 60S, 80S, monosome, and polysome fractions. Linear and circular RNAs were quantified by qRT-PCR and plotted by relevant quantity in each fraction. (B) Linear forms assayed were associated with monosome and polysome fractions. (C) Circular forms were predominantly unassociated with these complexes. Knockdown using three targeted siRNAs against HIPK3 or ZFY were quantified by qRT-PCR and plotted to show the effect of differentially targeted siRNA against the linear, circular, and both (lin/circ) forms of (D) HIPK3 and (E) ZFY. Quantities shown are given as ΔΔCt and are normalized to TBP and then to a nonspecific (NS) siRNA.