YY1 S180,184D phospho-mutant exhibits an increased DNA binding affinity in vitro.

<p>(<b>A</b>) Whole cell lysates were prepared from nococdazole treated HEK293 cells transiently transfected with Flag-vector, Flag-YY1 wild type, Flag-YY1 S180,184A and Flag-YY1 S180,184D followed by Western blot. The blot was probed with anti-pS184 antibody, then stripped and reprobed with anti-YY1 antibody. (<b>B</b>) Whole cell lysates were prepared from asynchronous HEK293 cells transiently transfected with Flag-vector, Flag-YY1 wild type, Flag-YY1 S180,184A and Flag-YY1 S180,184D. Cell lysates were used in an electrophoretic mobility shift assay (EMSA) using ³²P-labeled H3.2 α, P5-60 and p16 DNA oligonucleotides as probes. (<b>C</b>) The same lysates used in the EMSA were also used in a Western blot probed with anti-YY1 antibody to show equal Flag-YY1 levels.