FIGURE 2.

A pull‐down assay was performed with GST‐SIRT2 fusion protein from the lysate of B104 cells under control conditions (A) or treated with 10 ng/mL LPS for 12 h (B). Immunoblots showed Hsp90 interacts only with GST‐SIRT2, but not GST alone. LPS stimulation appears to promote binding of Hsp90 to SIRT2. Co‐immunoprecipitation was performed with anti‐Hsp90 (C and E) and anti‐Sirt2 (D and F). Immunoblots showed detection of SIRT2 in Hsp90 co‐IPs (C) and detection of Hsp90 in SIRT2 co‐IPs (D), suggesting that SIRT2 and Hsp90 form a complex in vivo. All assays were repeated at least three separate times yielding similar results, and representative immunoblots are shown. The precipitated protein complex was probed with anti‐Hsp90 (E) and anti‐SIRT2 (F) to confirm co‐IP with specific antibodies