Figure 2.

Effects of primer hybridization of AU-6 on g5p binding. (A) EMSA. ³²P-labeled AU-6 (1 μM) was heated at 95°C for 3 min in the absence (lanes 1–5) or presence (lanes 6–10) of 1.5 μM 3′P (3′ primer). The mixtures were cooled slowly at room temperature for 15 min to allow primer hybridization. Various concentrations (0–16 μM) of a DNA competitor (I-7c26) were added before adding 12.8 μM of g5p. The binding reaction was performed at 37°C for 30 min and subjected to a 2.5% agarose gel. Shown is a phosphor image of the dried gel. (B) Quantitation of complex formation inhibited by the competitor in the absence (open circle) or presence (solid circle) of 3′P. Percentages of g5p·AU-6 complexes (intermediate plus saturated) retained in each lane of (A) were quantitated and normalized to the samples without competitors (lane 1 or 6).