Figure 2

Effect of polyQ-expanded Atx3 on the function of P97 in down-regulating neddylation.
(a) Effect of polyQ-expanded Atx3 on the neddylation levels in cells. HEK 293T cells were co-transfected with HA-NEDD8 and FLAG-tagged Atx322Q, Atx3100Q or Atx3100Q/VBM*. About 72 hrs after transfection, the cell lysates were prepared for Western blotting with an anti-FLAG or anti-HA antibody. (b) Quantification of the neddylation levels by three Atx3 types. (c) Effect of polyQ-expanded Atx3-IC on the neddylation levels in cells. Same as (a), but the C-terminal fragments of Atx3 (residues 221–360; Atx322Q-IC, Atx3100Q-IC or Atx3100Q-IC/VBM*) applied. (d) Quantification of the neddylation levels by the three Atx3-IC forms. Data are shown as Means ± SEM (n = 4). *, p < 0.05; **, p < 0.01; ***, p < 0.001; N.S., no significance.