To determine whether eIF2 phosphorylation by carcinoma cell lysates is dsRNA dependent, the same assay was performed in the presence or absence of dsRNA (Figure 3B). Purified PKR from the transformed human embryonic kidney cell line, 293, gave rise to two radioactive bands of autophosphorylated PKR and phosphorylated eIF2 (lane 5). When the MCF7 mammary carcinoma cell lysate was assayed for eIF2 phosphorylation in the absence of dsRNA, eIF2 was phosphorylated to a very small extent (lane 4), which increased several fold in the presence of dsRNA (compare lanes 3, 4).
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