Fig. 1: NIX and BNIP3 are unable to bind FIP200 in vitro. | Nature Cell Biology

Fig. 1: NIX and BNIP3 are unable to bind FIP200 in vitro.

From: Reconstitution of BNIP3/NIX-mitophagy initiation reveals hierarchical flexibility of the autophagy machinery

Fig. 1

a, Schematic of the domain structures of NIX, BNIP3, FUNDC1 and BCL2L13. MER, minimal essential region; BH, Bcl-2 homology domain; TMD, transmembrane domain. b, Representative SDS–PAGE gels of NIX-GST, BNIP3-GST, FUNDC1-GST and BCL2L13-GST. Arrows indicate the predicted molecular weight. c–h, Microscopy-based bead assay of agarose beads coated with the indicated GST-tagged cargo receptors and incubated with GFP-tagged FIP200-CTR (residues 1429–1591) (c), GFP-tagged full-length FIP200 (d), FIP200-CTR and kinases TBK1, MBP-ULK1, CK2 or Src (Y530F; constitutively active mutant) (e), full-length FIP200 and kinases TBK1, MBP-ULK1, CK2 or Src (Y530F; constitutively active mutant) (f), FIP200-CTR and lambda protein phosphatase (g), full-length FIP200 and lambda protein phosphatase (h). Samples were analysed by confocal imaging. Scale bars, 100 µm. Results are representative of three replicates (b–h). Unprocessed blots are available in the source data. Schematic generated with BioRender.

Source data

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