Presence of HLTF in some of the DNA-protein complexes formed on the HS2-CS.
(A) In vitro sequence-specific binding of K562 nuclear proteins to the HS2-CS. The DNA probe used in the EMSA is 32P-labeled double-stranded HS2-CS sequence (oligo 10). The control lane is EMSA without nonradioactive HS2-CS oligo. The molar excesses of nonradioactive DNA probe used in the competitive EMSA are shown on the top of the lanes. The NS oligo is described in Figure 2. The EMSA bands are numbered and indicated by arrow marks. Free probe lane displays EMSA pattern in the absence of K562 nuclear extracts. (B) HLTF is present in 2 EMSA bands obtained with a double-stranded HS2-CS sequence and K562 nuclear extracts. Gel supershift/neutralization assay with K562 nuclear extract and 32P-labeled HS2-CS oligonucleotide in the presence (+) and absence (−) of HLTF antibody A and B, normal rabbit serum, and anti-Sin3A antibody. The assay conditions were the same as described in Figure 3.
