Figure 1.

Analysis of new interactions of Atg1 complex subunits by pull-down. (a) Protein extracts of Dictyostelium cells co-expressing GFP-Atg1 (or GFP as a control) and TKT fused with the HA epitope (TKT–HA) were subjected to pull-down with GFP trap and the interaction analysed by Western blot using anti-HA antibody. The same blot was then used for GFP and GFP-Atg1 detection with anti-GFP antibody. TKT–HA was pulled down by GFP-Atg1 and not by the GFP control. Note: due to the large amount of the GFP control, the signal was expanded to nearby lanes. (b) Similar experiment as in (a) to detect the interaction between GFP-Atg101 and AreA-HA. In this case, GFP-Atg101 and AreA-HA were expressed in different cells and their respective extracts mixed before performing pull-down experiments.