Degradation of endogenous Nurr1 in PC12 cells and ectopically expressed Nurr1 in HeLa cells.

<p>(A) PC12 cells were treated with CHX in the absence or in the presence of Lactacystin (Lacta) for the times indicated and cell extracts analyzed by immunoblot with anti-Nurr1 antibodies. (B, D and F) HeLa cells were transiently transfected with full-length Nurr1, N-terminal flag-tagged Nurr1 or Nurr1 1–337 as indicated, after transfection cells were treated with CHX in the absence or in the presence of Lactacystin (Lacta) for the times indicated and cell extracts analyzed by immunoblot with anti-Nurr1 antibodies (B and D). Protein loading control was assessed by immunobloting with anti-tubulin antibodies. (C and E) Graphs show the quantification of immunoblots, and results are expressed as means ± s. e. m. from three different experiments of the indicated Nurr1 protein constructs. F, transfected cells were analyzed by indirect immunofluorescence with anti-Nurr1 antibodies (red channel), counterstained for nuclei with DAPI (blue channel) and imaging by confocal microscopy for subcellular localization of Nurr1.</p>