Fig. 3: Electromobility Shift Assays (EMSA) with piggyBat transposon end sequences.

a pBat transposase shows multiple interactions in an EMSA DNA binding assay. LE44 with increasing concentrations of pBat-D237A. A single shifted species was observed with LE44 (lanes 1–7; 1: 0 nM protein; 2: 12.5 nM; 3: 25 nM; 4: 50 nM; 5: 100 nM; 6: 200 nM; 7: 300 nM) but not with a random oligonucleotide of the same length (lanes 8–14; protein concentrations are the same as lanes 1–7). b LE88 with increasing concentrations of pBat-D237A. In this case, two major shifted species were observed with LE88 (lanes 1–7; protein concentrations as above); these were not observed with a random oligonucleotide of the same length (lanes 8–14; protein concentrations as above). The red and green asterisks indicate the color of the fluorescent label. c The effect on pBat-LE88 binding by scrambling (“scr”) the sequence of either the first 44 bp of LE88 (“scr1-44LE45-88”) or (d) bp 45–88 (“LE44scr45-88”). e pBat binding to 100 nM RE100. Left to right, lanes correspond to 0 nM protein, 50, 100, 200, 400, and 600 nM. Three shifted species were observed (indicated by arrows). f pBat binds independently to oligonucleotides containing the three repeated pairs of motifs on the Left End (LE44, LE45-88, and LE89-132). All experiments were performed at least two times with similar results. Source data are provided as a Source Data file.