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. 2016 Jun 6;17(7):1044–1060. doi: 10.15252/embr.201541960

Figure EV4. Atg19 solubilization effect on prApe1 dodecamers (related to Fig 3).

Figure EV4

  1. Size‐exclusion chromatography profile (Superose 6 column) of Atg19 after anti‐MBP‐Atg19 purification and cleavage of the MBP tag (left). Shown in gray is the overlay with the profile of the prApe1/Atg19 complex after anti‐MBP‐Atg19 purification and cleavage of the MBP tag. SDS–PAGE of peak fractions of the gel filtration profile of Atg19 (right).
  2. Size‐exclusion chromatography profile (Superose 6 column) of prApe1 after anti‐GST‐prApe1 purification and cleavage of the GST tag (left). Shown in gray is the overlay with the profile of the prApe1/Atg19 complex after anti‐MBP‐Atg19 purification and cleavage of the MBP tag. SDS–PAGE of peak fractions of the gel filtration profile of prApe1 (right). Samples were run on three separate gels.
  3. Representative SDS–PAGE of pelletation assay of purified prApe1 dodecamers alone, purified Atg19 alone, and the purified prApe1/Atg19 complex.
  4. Bar plot of gel band intensities from supernatant and pellet fraction of (C).
  5. Western blot of prApe1 (top) and Atg19 (bottom) showing clarified lysate from prApe1‐GFP/Atg19‐mCherry/ypt7Δ (lane 1) and prApe1‐GFP/Atg19Δ/ypt7Δ (lane 2) Saccharomyces cerevisiae cells.
Data information: Data in (C) and (D) are representative of three independent experiments. Error bars indicate SD. Analysis of variance (ANOVA) was performed to assess whether the supernatant to pellet ratio of prApe1 changed upon the presence of Atg19 (D): ***P = 1.7 × 10−4.