Extended Data Fig. 6: Degradation of a neo-cargo by selective autophagy.

a, Indicated cells expressing Atg19 or Cnb1–Atg19, and GFP–Ape1 and copper-inducible untagged Ape1 or FKBP–GFP-µNS were grown to mid-log phase in the presence of 50 µM CuSO₄ and treated with FK506. GFP cleavage was monitored by TCA precipitation and immunoblotting. One out of three biological replicates with similar results is shown. b, atg19∆ cells expressing Atg19, an empty vector control (–), Cnb1–Atg19 or Atg19–GBP were grown to mid-log phase. Cell extracts were analysed by immunoblotting. One out of three biological replicates with similar results is shown. c, GFP–Atg11 atg19∆ cells expressing endogenous BFP–Ape1 and copper-inducible untagged Ape1 and GFP–Atg11 atg19∆ ape1∆ cells expressing pp-BFP–µNS or Atg19-BFP–µNS were grown to mid-log phase. GFP–Atg11 structures were photobleached, and recovery of the signal was monitored. Scale bar: 1 µm. Quantification: recovery of the GFP signal. Data are mean values ± SEM (n > 26 structures per condition across replicates, three biological replicates). d, Cells from Extended Data Fig. 6a coexpressing mScarlet–Atg11 were analysed and quantified as in Fig. 2b. Scale bar: 2 µm. Quantification: GFP clustering as the coefficient of variance (SD/mean GFP intensity) in a box plot. Horizontal lines: median, box: 25th to 75th percentiles, whiskers: expand to 5th and 95th percentiles, circles: mean value of each replicate, outliers: black dots (n = 50 structures per condition and replicate, three biological replicates). One-tailed unpaired t-test. P value: P < 0.0001. For each panel, one out of three biological replicates is shown. Source numerical data and unprocessed blots are available in source data, arbitrary units (a.u.).

Source data