Actinomycin D induces the
formation of complex B and C with TNFα mRNA.
Probe TNF-10 was incubated alone (lane 1) or with protein extracts prepared
from B10R cells untreated (lanes 2, 4, 6 and 8) or treated with
5 µg/ml of Actinomycin D for
1 h (lanes 3, 5, 7 and 9). Lanes 2 and 3, total cellular extract; lanes
4 and 5, nuclear wash fraction; lanes 6 and 7, nucleoplasm; lanes
8 and 9, total cytoplasmic extract. The resulting RNA–protein
complexes were analyzed either on the non-denaturing gel (A)
or UV-crosslinked and analyzed on denaturing 8% polyacrylamide–SDS
gel (B).