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. 2000 Jun 20;97(14):7841–7846. doi: 10.1073/pnas.140202897

Figure 3.

Figure 3

Association of chaperone proteins with HD exon 1 in vitro. (A) Schematic representation of the structures of the polyQ-containing GST fusion proteins analyzed. (B) DnaJ or DnaK (6 μM each) were incubated with uncleaved GST-HD20Q, GST-HD53Q, GST-HD53QΔP, or GST-MJD35Q (3 μM each) and then precipitated with glutathione-Sepharose. GST-HD fusion proteins and associated proteins were eluted with reduced glutathione (lanes 1–6 and 8–13) or Mg-ATP (lanes 7 and 14), followed by SDS/PAGE and immunoblotting with anti-DnaJ (lanes 1–7) or anti-DnaK antibodies (lanes 8–14). Chaperones were added to Sepharose beads alone (lanes 1 and 8) or to beads containing GST (lanes 2 and 9), GST-HD20Q (lanes 3 and 10), GST-HD53Q (lanes 4, 7, 11, and 14), GST-HD53QΔP (lanes 5 and 12), or GST-MJD35Q (lanes 6 and 13).