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. 2009 Apr 8;4(4):e5118. doi: 10.1371/journal.pone.0005118

Figure 2. No physical interaction between HP1 proteins and cohesin.

Figure 2

(A) Immunoprecipitation of native proteins from HeLa nuclear extracts with non-immune rabbit IgG (lane 2) or mouse preimmune serum (lane 4) and specific antibodies against cohesin SMC3 subunit (lane 3), HP1alpha (lane 5), HP1beta (lane 6) and HP1gamma (lane 7). The immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. An aliquot of the extract was also loaded (lane 1). (B) Immunoprecipitation reactions (IP) carried out with non-immune mouse IgG (control) or anti-HP1gamma from the soluble (lanes 3–4) and chromatin-bound fractions (lanes 7–8) of HeLa cells treated with 1% paraformaldehyde were analyzed by immunoblotting. Aliquots of the corresponding flowthroughs (FT, lanes 1–2 and 5–6) were also loaded. (C) Aliquots of a HeLa cell nuclear extract were incubated with GST agarose bound to GST, HP1alpha-GST, HP1beta-GST or HP1gamma-GST fusion proteins (lanes 2–5, respectively). After washing, the agarose-bound proteins were analyzed by immunoblotting. An aliquot of the HeLa cell extract was also analyzed (lane 1).