Figure 2

A, electrophoretic mobility shift assay (EMSA) with radiolabeled either -1952T or -1952C probe and HEK-293 cell nuclear extracts (NE) and B, EMSA assays with radiolabeled either -2012T/-2012C and +561G/+561A probes and HEK-293 cell NE. A. Lanes 1 and 6, mobility of the labelled probes without NE; lanes 2 and 7, mobility of the labelled probes with NE in the absence of competitor. A specific nuclear protein binding can be almost completely abolished both by 100-fold unlabeled -1952T but not with -1952C probe (lanes 4 and 5; lanes 9 and 10). Super shift assays incubating with anti-HSF1 antibody did not show any super shifted protein complex (lanes 3 and 8). B. Lanes 5, 10, 15 and 20, mobility of the labelled probes without NE; lanes 4, 9, 14, and 19, mobility of the labelled probes with NE in the absence of competitor. Competition assays with 100-fold unlabeled probes (lanes 1 and 2; lanes 6 and 7; lanes 11 and 12; lanes 16 and 17). Super shift assays incubating with anti-HSF1 antibody did not show any super shifted protein complex (lanes 3, 8, 13 and 18).