Association of Htt-552 and exogenous CMA component proteins.

<p>(A) Exogenous expressed LAMP-2A or Hsc70 co-localized with exogenous Htt-552-18Q and Htt-552-100Q in HeLa cells. Cells were transfected with Htt-552 for 48 h and fixed with methanol, blocked, and processed for double immunofluorescence with antibodies against LAMP-2A (green), Hsc70 (green) and Htt-552 (red). Merged images of both channels are shown at the bottom. N: nucleus. Thin arrows point to Htt-552 immunoreactivity. Thick arrows point to lysosomes. The scale bar = 10 µm. (B) Co-immunoprecipitation of exogenous expressed LAMP-2A with exogenous Htt-552. Upper panel: PC12 cell were transfected with PCDNA4-LAMP-2A for 72 h. Cell lysates were subjected to immunoprecipitation with anti-Htt antibody or no primary antibody and Western blot analysis was performed with anti-LAMP-2A antibody. Lower panel: PC12 cell were transfected with Htt-552 for 48 h. Cell lysates were subjected to immunoprecipitation with anti-LAMP-2A antibody or no primary antibody and Western blot analysis was performed with anti-Htt antibody. (C) Co-immunoprecipitation of exogenous expressed Hsc70 with Htt-552 from PC12 cells. Upper panel: PC12 cell were transfected with PCDNA4-Hsc70 for 72 h. Cell lysates were subjected to immunoprecipitation with anti-Hsc70 antibody or no primary antibody and Western blot analysis was performed with anti-Htt antibody. Lower panels:PC12 cell were transfected with Htt-552 for 48 h. Cell lysates were subjected to immunoprecipitation with anti-Htt antibody or no primary antibody and Western blot analysis was performed with anti-Hsc70 antibody. Ab: Antibody.