Figure 2.

Electrophoretic mobility shift assay of NF-Y binding to Dmrt7 promoter. The oligo1 corresponding to -58/-31 and oligo2 corresponding to -20/+8 were γ-32P-ATP labelled and incubated with 5µg of nuclear extract of mouse testis in the absence or presence of 50-fold excess various competitor DNA (mutant or non-labeled oligo) or anti-NFYa antibody, as indicated on the top of the gel image. Samples were resolved on 5% polyacrylamide gels in 0.5% TBE running buffer at 10 V/cm for 2 h. The specific DNA/protein complex and the super-shift bands were indicated by arrows. The shift bands cannot be competed with mutated DNA competitor (lane 7, 8). The sequences of oligo1, oligo2 and corresponding mutants are shown in the lower panel.