Skip to main content
Figure 7 | Journal of Molecular Signaling

Figure 7

From: Defining regulatory and phosphoinositide-binding sites in the human WIPI-1 β-propeller responsible for autophagosomal membrane localization downstream of mTORC1 inhibition

Figure 7

Protein-phospholipid overlay assays with wild-type and mutant GFP-WIPI-1 variants. From U2OS cells transiently expressing GFP-WIPI-1 or GFP-tagged mutant WIPI-1 variants native cell extracts were generated in parallel and used to overlay membrane-immobilized PtdIns(3)P (12,5–200 pmol) followed by anti-GFP ECL detection. Prior to overlaying the membranes with native cell extracts, the volumes of the different extracts were adjusted to include equivalent levels of GFP-WIPI-1 wild-type or mutant protein, judged by anti-GFP western blotting (not shown). Representative results are shown (n = 3). In red, GFP-tagged WIPI-1 mutants unable to bind to PtdIns(3)P.

Back to article page