Figure 4.

ATXN3 directly interacts with GABARAP and LC3C. (a) Purified, recombinant ATXN3 was incubated with blocked agarose beads (empty beads, e.b.) or recombinant GABARAP/LC3A immobilized on agarose beads. Binding was analyzed by immunoblotting. (b) Cell lysates from HeLa cells expressing doxycycline (dox)‐inducible ^10xHisATXN3^HA were incubated with blocked agarose beads (e.b.) or recombinant GABARAP/LC3A immobilized on agarose beads. Binding of endogenous ATXN3 and ^10xHisATXN3^HA was analyzed by immunoblotting with the indicated antibodies; * indicates band corresponding to ATXN3 from previous detection. (c) Cell lysates from HeLa cells expressing dox‐inducible ^10xHisATXN3^HA were incubated with purified, recombinant GST‐tagged Atg8 proteins and binding analyzed by immunoblotting with the indicated antibodies. (d) Schematic illustration of GFP‐ATXN3 constructs used in (e). (e) Cell lysates from HeLa cells transfected with the indicated constructs were incubated with blocked empty agarose beads (e.b.) or recombinant GABARAP immobilized on agarose beads. Binding was analyzed with immunoblotting using the indicated antibodies