Fig. 7.
The activities of per unit protein A recruiting (+)RNA1E and self-interaction are regulated by manipulation of phospholipids in cells. (A) Pr-E cells expressing (+)RNA1E together with protein AGAA-His plus empty vector-HA (lane 1) or protein AGAA-His plus protein AGAA-HA (lanes 2–6) were harvested. Cell Lysates were immunoprecipitated with an anti-His antibody (lanes 1 and 3–6) or control IgG (lane 2). The immunoprecipitated complexes were divided into two equal fractions. One of fractions was blotted with an anti-HA antibody to determine the self-interaction of protein A. The (+)RNA1E associated with the immunoprecipitated protein A was separated from the other fraction and following analyzed by real-time RT-PCR. (B) Graph of the ratios of the accumulation of (+)RNA1E analyzed by real-time RT-PCR as described above and protein A self-interaction versus protein A׳s accumulation. The accumulation of RNA and protein is normalized to 18S rRNA and GAPDH from total cells, respectively. Error bars represent S.D. values from at least three independent experiments.