Figure 4 | Scientific Reports

Figure 4

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

Figure 4

Co-precipitation of diUb and P97 by Atx3 as detected by fractionation and SDS-PAGE analysis.

(a) K48-diUb by Atx322Q-IIC. (b) K48-diUb by Atx346Q-IIC. (c) P97-ND1 by Atx322Q-IIC. (d) P97-ND1 by Atx346Q-IIC. The protein mixtures were centrifuged to separate supernatant (Sup.) and pellet (Pel.) and analyzed by SDS-PAGE (15% gel). The stock concentrations of Atx322Q-IIC and Atx346Q-IIC were 600 μM. Atx322Q-IIC or Atx346Q-IIC was mixed with K48-diUb (60 μM) at an increased molar ratio of Atx3-IIC/K48-diUb (0:1, 1.5:1, 3:1 and 6:1), while it was mixed with P97-ND1 (30 μM) at an increased molar ratio (0:1, 3:1, 6:1 and 12:1). Atx3-IIC, C-terminal fragment (residues 221–373) of isoform-II Atx3. K48-diUb, K48-linked diubiquitin.

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