Figure 1 | Scientific Reports

Figure 1

From: Aggregation of polyglutamine-expanded ataxin-3 sequesters its specific interacting partners into inclusions: Implication in a loss-of-function pathology

Figure 1

Sequestration of P97 into insoluble aggregates by Atx3.

(a) Domain architecture of the two isoforms of Atx3. Josephin domain, a deubiquitinating domain of Atx3; UIM, ubiquitin-interacting motif; VBM, VCP-binding motif; HR, hydrophobic region; Qn, polyQ tract. The expanded glutamine residues are not included in the numbering. (b) Sequestration of overexpressed P97. FLAG-tagged Atx322Q, Atx3100Q or Atx3100Q/VBM* (282RKRR to HNHH) was co-transfected with HA-P97 into HEK 293T cells. The cell lysates were subjected to fractionation and Western blotting with an anti-FLAG or anti-HA antibody. The constructs contain sequences encoding the amino-acid residues of 1 - 360. Sup., supernatant; Pel., pellet. (c) Quantification of the amounts of overexpressed HA-P97 in supernatant and pellet fractions. (d) Sequestration of endogenous P97. FLAG-tagged Atx322Q, Atx3100Q or Atx3100Q/VBM* was transfected into HEK 293T cells. The cell lysates were subjected to fractionation and Western blotting with an antibody against P97. (e) Quantification of the amounts of endogenous P97 in supernatant and pellet fractions. Data are shown as Means ± SEM (n = 3). *, p < 0.05; **, p < 0.01; ***, p < 0.001; N.S., no significance.

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