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. Author manuscript; available in PMC: 2012 Jun 26.
Published in final edited form as: Mol Cell. 2011 Jun 24;42(6):758–770. doi: 10.1016/j.molcel.2011.05.010

Figure 2. Trc35 maintains a cytosolic pool of Bag6.

Figure 2

A, Trc35 expression alters the subcellular localization of endogenous Bag6. HeLa cells transfected (T) with a control vector (panels 1–3) or a Trc35-FLAG expressing plasmid were stained with anti-Bag6 (red) and -FLAG (green) antibodies. Where indicated, DNA was stained with DAPI (blue). B, Trc35, but not Ubl4A, retains overexpressed Bag6 in the cytoplasm. HeLa cells transfected with the indicated plasmids either individually (panels 1–3) or in combination (panels 4–9) were stained to visualize Bag6 (red), FLAG-tagged proteins (green), and DNA (blue). C, D, Knockdown of Trc35 increases nuclear localization of Bag6. HeLa cells transfected with either a control or Trc35 shRNA construct were stained with a Bag6 antibody (red) and DAPI (blue). Shown are representative cells imaged by a confocal microscope. The graphs in D show the profiles of averaged Bag6 (green) and DAPI (blue) fluorescence intensity across the nuclear envelope boundary (indicated by position 0) measured by a confocal microscope (n=10 cells). See also Figure S2