Extended Data Fig. 8: Loss of recycling by spe-39(RNAi) or vps-45(RNAi) partially rescues a sand-1 mutant and endogenous VIPAS39 interacts with SNX1.
From: FERARI is required for Rab11-dependent endocytic recycling
a, Knock-down of spe-39 or vps-45 induces the degradation pathway in coelomocytes. In sand-1(ok1963) coelomocytes BSA-TR is not transported to the lysosomes (LMP-1 compartment). spe-39(RNAi) and vps-45(RNAi) will favor the transport of BSA-TR to coelomocytes, probably by abolishing recycling. Arrows indicate LMP-1-positive compartments containing BSA-TR. n=3 independent RNAi experiments (n=20 animals). Scale bars:5 µm. b, GFP-RME-1 levels were unchanged upon FERARI (RNAi). Western blot analysis of GFP-RME-1 levels in FERARI(RNAi) worm lysates. n=3 independent experiments. c, Endogenous VIPAS39 does not interact with Retromer subunit VPS35 in HEK-293 cells transfected with GFP-VPS35. n=3 independent experiments. d, GFP-SNX1 transfected HEK-293 cells show slight interaction between SNX1 and endogenous VIPAS39. n=3 independent experiments. Very weak interaction (<1%) could not be measured due to overexposed bands in the input. See Unprocessed Blots Extended Data 8.
