Fig. 2
From: Kaiso mediates human ICR1 methylation maintenance and H19 transcriptional fine regulation
Kaiso binds the ICR1-B4-KBS in vitro. EMSA was carried out with fluorescence-labelled oligonucleotide probes for the ICR1-B4-KBS (lanes 1–8). N-terminal Flag-tagged Kaiso protein (in vitro FLAG-Kaiso) was produced by in vitro translation, using modified HeLa cell extracts (Pierce). The ds-oligo for the ICR1-B4-KBS (lane 1) is shifted by in vitro FLAG-Kaiso—Kaiso (lanes 3 and 5), but not by the cell extract containing the expression vector alone (neg. control, lane 2). Complexes were competed by an excess of unlabelled ICR1-B4-KBS ds-oligo (lane 4) but not with a non-specific control oligo (lane 5). In supershift reactions, Flag-tagged Kaiso was incubated with anti-Kaiso (lane 6), anti-WT1 (lane 7) or anti-Flag (lane 8) antibodies, the anti-WT1 representing a non-specific control