Fig. 3: Pho81 functionally interacts with Atg13 and Atg17 during pexophagy upon P-S.
From: A metabolite sensor subunit of the Atg1/ULK complex regulates selective autophagy
a, Y2H assay of cells expressing pGAD-PHO81 or pGAD-ATG36 in combination with pGBDU (empty vector, ev), pGBDU-ATG8 or pGBDU-ATG11 grown on SD + HIS or SD − HIS medium. b–d, Pex11–GFP turnover in indicated strains after P-S (24 h) (b, n = 3–6; c, n = 9 biologically independent experiments) or after N-S (24 h) (n = 4 biologically independent experiments) (d). Data are mean ± s.d. e, 2GFP–Atg8 turnover in indicated strains after P-S (24 h). Data are mean ± s.d. (n = 6 biologically independent experiments). f, 2GFP turnover in indicated strains after P-S (24 h). Data are mean ± s.d. (n = 3 biologically independent experiments). g, Fluorescence imaging of WT cells expressing genomic PHO81–GFP and ATG13–mCherry after P-S (4 h). Arrowheads indicate Pho81–GFP positive (white) or negative (purple) Atg13 foci. Scale bar, 5 µm. Quantification of Atg13–mCherry and Pho81–GFP co-localization in WT cells. Data are mean ± s.d. (n = 150 cells examined over three independent experiments). h, Fluorescence imaging of indicated strains expressing genomic PHO81–GFP and ATG1–mCherry after P-S (4 h). Arrowheads indicate Pho81–GFP-positive (white) or Pho81–GFP-negative (purple) Atg1 puncta. Scale bar, 5 µm. Quantification of Atg1–mCherry and Pho81–GFP co-localization in WT and ∆atg13 cells. Data are mean ± s.d. (n = 300 cells examined over three independent experiments). Statistical significance was assessed using one-way ANOVA followed by Tukey’s multiple comparisons test. P values relative to WT or as indicated: ***P < 0.0001 except *P = 0.0494 (d) and P values relative to ∆pho81 ###, P < 0.0001 (b–f). Two-way ANOVA followed by Tukey’s multiple comparisons test. P values relative to WT or as indicated: ***P = 0.0002 (g), **P = 0.0078 (h), non-significant (n.s.). Source numerical data and unprocessed blots are available in Source data.
