Figure 3.

RNA Immunoprecipitation Assays to Demonstrate the Interaction between TFIIIA and PSTVd in Vivo.

The cell lysates were incubated with anti-HA antibodies (α-HA) to detect HA-tagged TFIIIA-7ZF and -9ZF proteins or with anti-myc antibodies (α-myc) as a negative control for specificity. The immunoprecipitated fractions were subject to PSTVd detection by RT-PCR. 5S rRNA was used as a positive control while U6 snRNA was used as a negative control. The presence of both the (+)- and (−)-PSTVd RNAs was examined in the immunoprecipitate. The quantitation and the statistical analyses are shown at the bottom panel. Briefly, the signals from the input lysates served as a reference representing 1% for RNA and 10% for protein for standardization. The band intensity in negative (α-myc) and positive (α- HA) treatments was quantitated and normalized to the signals of inputs to obtain their percentage values. Three biological repeats were used for the two-tailed t test (**P < 0.01 and *P < 0.05).