Supplementary Figure 3: Mutational analyses of ScAtg2.
From: Atg2 mediates direct lipid transfer between membranes for autophagosome formation
a, SDS-PAGE of purified recombinant proteins used for in vitro analyses. Protein bands were stained with Coomassie Brilliant Blue. Uncropped gel images are shown in Supplementary Data Set 1. b, The effect of ATP on phospholipid transfer mediated by Atg2. c, Comparison of the MT activity between Atg2 and the Vps13 fragment analyzed by dynamic light scattering. d, MT activities of Atg2 as assessed by fluorescence microscopy of NBD-PE and Rhod-PE liposomes. Scale bar indicates 5 μm. e, Bulk autophagic activity measured by the ALP assay. In e and f, data are presented as mean±s.d. from n = 3 independent experiments. N.S. means not statistically significant (two-tailed Student’s test). f, Maturation of a selective autophagy cargo aminopeptidase I (Ape1) via autophagy monitored by western blotting. Uncropped blot image is shown in Supplementary Data Set 1. g, LT and MT activities of the K108E R216E mutant. h, Confocal microscopic analysis of liposomes mixed with the 3D mutant of ScAtg2. i, Circular dichroism spectra of wild-type and 3D mutant of MBP-fused SpAtg2NR.
