Fig. 7.

Synthesis of nNOS protein in vitrois suppressed by the antisense pseudo-NOS RNA. Lane 1represents the result of the translation of NOS cRNA and shows the main labeled product is a protein of the expected (93 kDa) size (arrow). Lane 2 illustrates the effect of incubating the 0.2 μg of NOS cRNA with 2 μg of antisense pseudo-NOS cRNA before translation. Note strong suppression of translation of the nNOS protein. As a control for any effects on translation that are not related to the formation of a duplex, we also preincubated the 0.2 μg of NOS cRNA with a 2 μg of a sense version of the pseudo-NOS cRNA (lane 3). No inhibition of NOS protein synthesis is observed. Similar results, although weaker, have been obtained, even when the ratio between nNOS cRNA and pseudo-NOS transcripts was 2.5:1. A second major protein of ∼50 kDa can be seen in eachlane. This protein is present in the cell-free wheat germ system and performs a useful function as an internal control. Note that it is not diminished in intensity in lane 2.