Rab GTPases interact with cargo receptors for autophagic degradation. (A) The interaction of Rab GTPases with selective autophagy receptors. Flag-tagged selective autophagy receptors were expressed individually in HEK293T cells, which simultaneously expressed HA-GFP-Rab GTPases. IP was performed with anti-HA beads, which was followed by western blot for Flag-tagged selective autophagy receptors. (B) Summary of interactions between Rab GTPases and selective autophagy receptors. (C) In vitro interaction between GST-NDP52 WT, GST-NDP52 mutants, and the Rab GTPases were analyzed by GST pulldown assay using purified recombinant proteins (in Fig. S2, E and F). (D) Illustration of NDP52 domain interacting with Rab GTPases. (E) Recombinant protein of FLAG-NDP52 was incubated with immobilized GST-Rab8 loaded with GDP or GTP-γ-S for GST pull-down assay. The beads were washed and analyzed by western blot for FLAG (NDP52). The amounts of GST proteins were indicated by Coomassie blue staining. Molecular weight measurements are in kD. Source data are available for this figure: SourceData F4.