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. 2003 Mar;9(3):331–345. doi: 10.1261/rna.2144203

FIGURE 3.

FIGURE 3.

Transient transfection of En5-3 cells with Ntat2ANeo(SI) and related mutants containing nucleotide substitutions within the putative loop segment of the SL1 stem-loop located at the 3′ end of the 3′NTR. (A) Schematic showing the substitution mutations within the loop segment. Nucleotides are numbered according to their position from the 3′ end of the RNA. (B) SEAP activity present in supernatant culture fluids collected at 24-h intervals following transfection of En5-3 cells with Ntat2ANeo(SI) (filled circles), BLoop, in which the entire loop sequence was replaced with its complement (filled squares), or mutants with individual nucleotide substitutions: 21A (filled triangles), 22G (asterisks), 23U (open diamonds), 24A (open circles), 25U (open triangles) or 26C (dash). Results are also shown following transfection of Ntat2ANeo(SI) containing the ΔGDD mutation that is lethal for replication (filled diamond). The transfected cells were passaged after collection of the supernatant fluid at day 7 (open arrow).