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. 2002 Dec;22(23):8114–8121. doi: 10.1128/MCB.22.23.8114-8121.2002

FIG. 3.

FIG. 3.

Decapping assays with immunopurified and recombinant hDcp1a and hDcp2. α-32P-m7cap-labeled RNA was incubated with immunopurified FLAG-tagged hDcp1a (lanes 4, 5, 9, and 10; corresponding to 0.2 × 106, 1 × 106, 1 × 106, and 1 × 106 cells' worth of protein, respectively), hDcp2 (lanes 6 and 7; 1 × 106 and 5 × 106 cells' worth, respectively), mock-purified cell extract (lanes 2 and 3; 1 × 106 and 5 × 106 cells' worth, respectively), or no cell extract (lanes 1, 8, and 11) or with purified bacterially expressed GST (lanes 12 and 13; 10 and 100 ng, respectively), GST-hDcp1a (lanes 14 and 15; 10 and 100 ng, respectively), GST-hDcp2 (lanes 16 and 17; 10 and 100 ng, respectively), or GST-hDcp1a and GST-hDcp2 (lanes 18 and 19; 10 and 100 ng each, respectively), and products were separated by thin-layer chromatography. The reaction mixture in lane 10 was treated with nucleoside diphosphate kinase (NDPK) and ATP. The migration of unlabeled GMP, GDP, m7GMP, m7GDP, and m7GTP (25 μg) is indicated on the left.