CTGC DNA motifs are essential for the stable formation of PICH-1, -2 and −3 on TASHET in vitro . EMSAs were performed as detailed in Figure 2. (A) Mutations used in EMSA and transient transfections shown in Figures 7 and 8: The name of the mutation is indicated at the left of the sequence. The mutated nucleotides are indicated in white. The purple box indicates the TATA box. The blue boxes indicate the CTGC motifs: the CTGC5’ motif is located just upstream of the TATA box, and two CTGC3’ motifs are located just downstream of the TATA box, the first one in the reverse orientation, the second one in the forward orientation. The mutations are named after the displayed mutations. (B) The indicated unlabelled competitors were added to the reaction (lanes 2 to 5). Lane 1 contains no competitor. (C) Relative intensity of the different complexes was calculated by phosphorimager analysis of the EMSA shown in B. Results are expressed relative to the intensity observed without competitor that was set to 1 for each complex.