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. 2007 Feb 5;27(7):2661–2675. doi: 10.1128/MCB.01098-06

FIG. 8.

FIG. 8.

wt HIC1 and the K314R mutant interact with SIRT1 and HDAC4. (A) Cos-7 cells were mock transfected (lane 1) or transfected with expression vectors for FLAG-tagged HIC1 proteins (1.25 μg; wild type or K314R mutant) and SIRT1 (1.25 μg) (lanes 2 to 6). HIC1 proteins were immunoprecipitated (IP) from cell lysates with anti-HIC1 2563 polyclonal antibodies. The resulting immunoprecipitates were then Western blotted and analyzed with the anti-SIRT1 monoclonal antibody (upper panel). The blot was stripped and probed with the rabbit anti-HIC1 polyclonal antibody (325) to ascertain the presence of HIC1 (middle panel). Two percent of each total cell extract (Input) was resolved by SDS-PAGE and immunoblotted with the anti-SIRT1 antibody (lower panel). The asterisk refers to a nonspecific band as specified by the supplier (Upstate). (B) A similar experiment was conducted as described above but with expression vectors for FLAG-HIC1 proteins (wild type or K314R mutant) and HA-tagged HDAC4. HDAC4 proteins were detected by using an anti-HA monoclonal antibody (Babco). −, absence of; +, presence of.

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