Fig. 5: WIPI2 and WIPI3 bind directly to the ULK1 complex.
a, Mitophagy flux measured by flow cytometry in WT HeLa cells expressing Fis1-FRB, FKBP–GFP–WIPI2 WT or ATG16L1-binding mutant R108E/R125E, and mt-mKeima, not induced or induced for 24 h by rapalog treatment. Data are presented as mean ± s.d. (n = 5 biologically independent experiments). Two-way ANOVA with Šídák’s multiple comparisons test. b, Microscopy-based bead assay of agarose beads coated with GFP-tagged ULK1 complex (consisting of FIP200-GFP, ULK1, ATG13, ATG101) and incubated with mCherry-tagged WIPI proteins. c, As in b, but with GFP-tagged ATG13/101 subcomplex and incubated with mCherry-tagged WIPI proteins. d, As in b, but with GFP-tagged FIP200-coated beads and incubated with mCherry-tagged WIPI proteins. e, As in b, but with GFP-tagged ULK1-coated beads and incubated with mCherry-tagged WIPI proteins. f, As in b, but with GFP-tagged ATG13/101-coated agarose beads incubated with mCherry-tagged full-length (FL) or IDR only (residues 364–425) WIPI2d. g, Mitophagy flux measured by flow cytometry in WT HeLa cells expressing Fis1-FRB, FL or IDR only (364–425 aa) FKBP–GFP–WIPI2, and mt-mKeima, not induced or induced for 24 h by rapalog treatment. Data are presented as mean ± s.d. (n = 4 biologically independent experiments). Two-way ANOVA with Šídák’s multiple comparisons test. ****P < 0.0001. NS, not significant. Results are representative of three biologically independent replicates (b–f). Scale bars, 100 µm. Source numerical data, including exact P values, are available in the source data.
