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. 2010 Jan 6;285(10):7827–7837. doi: 10.1074/jbc.M109.040618

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Hsp70 interacts with TRIM5α. A, the subcellular localization of endogenous Hsp70 and HA-tagged rhesus TRIM5α was evaluated by double labeling with anti-Hsp70 (red) and anti-HA (green) antibodies in HeLa cells stably expressing HA-tagged rhesus TRIM5α. B, co-immunoprecipitation of FLAG-Hsp70 with HA-TRIM5α from the cell lysates of HEK293T cells transfected with FLAG-Hsp70 and HA-tagged rhesus TRIM5α. Immunoprecipitates of FLAG antibody or HA antibody were analyzed for HA or FLAG by Western blotting. Total cell lysates were also analyzed for HA-TRIM5α, FLAG-Hsp70, or β-actin. C, co-immunoprecipitation of endogenous Hsp70 with HA-TRIM5α from the cell lysates of HeLa cells stably expressing HA-tagged rhesus TRIM5α or AGM TRIM5α. Immunoprecipitates of HA antibody were analyzed for Hsp70 by Western blotting. The arrow indicates the band of IgG heavy chain (labeled as H.C.). Total cell lysates were also analyzed for HA-TRIM5α, Hsp70, or β-actin. D, HEK293T cells were transfected with HA-tagged rhesus TRIM5α and FLAG-tagged Hsp70, Hsc70, or Hsp90, and the cell lysates were subjected to immunoprecipitation and Western blotting. Immunoprecipitates of HA antibody were analyzed for FLAG-Hsp and HA-TRIM5α. Total cell lysates were also analyzed for FLAG-Hsp and β-actin.