Fig. 3: Direct interaction between ULK1C and PI3KC3-C1.
From: Structure and activation of the human autophagy-initiating ULK1C:PI3KC3-C1 supercomplex
a, Bead-binding assay of ULK1C core and PI3KC3-C1. GSH beads coated with GST tag alone or GST-tagged PI3KC3-C1 was used to recruit various combination of GFP-tagged proteins. The schematic drawing illustrates the bait and prey samples used for each condition. b, Complementary bead-binding assay for confirming the interaction between FIP200 and PI3KC3-C1. GST-tagged various proteins were used to recruit the mCherry-tagged PI3KC3-C1. c, SDS–PAGE of the ULK1C:PI3KC3-C1 pulldown sample. Strep resin was used to pull down the TSF-tagged VPS15. The eluate was obtained by washing the resin with buffer containing 10 mM d-desthiobiotin and visualized by SDS–PAGE and Coomassie blue staining. The experiment was repeated two times with similar results.
