Skip to main content
. 2016 Jan 7;17(2):202–219. doi: 10.15252/embr.201540795

Figure 4. Formation of the exomer tetramer is dependent on ChAP concentration.

Figure 4

  • A, B
    Overexpression of Bch2 in absence of other ChAPs results in more efficient Chs5 recruitment and stabilization at the trans‐Golgi network (TGN). Fluorescence images of cells expressing Chs5‐GFP (A) and binding kinetics of Chs5‐GFP at the TGN (B) in WT, BCH2 ΔΔΔ, and GPD‐BCH2 ΔΔΔ strains. Scale bar, 5 μm. The mean of 20–30 FRAP measurements from different cells is shown. Calculated parameters are shown in Table 2. The schematic cartoons depict the predicted majority of the analyzed complexes: Chs5 dimer, violet ovals; Bch2, dark blue circles.
  • C, D
    Overexpression of Chs6 in absence of other ChAPs results in more efficient Chs5 recruitment and stabilization at the TGN. (C) Fluorescence images of cells expressing Chs5‐GFP in WT, CHS6 ΔΔΔ, and GPD‐CHS6 ΔΔΔ strains. Scale bar, 5 μm. (D) Binding kinetics of Chs5‐GFP at the TGN in WT, ΔΔΔΔ, CHS6 ΔΔΔ, and GPD‐CHS6 ΔΔΔ strains. Data processing was carried out as in (A and B). The schematic cartoons depict the predicted majority of the analyzed complexes; Chs5 dimer: violet ovals, Chs6: light blue circles.
  • E–G
    Overexpression of Chs6 rescues Chs3 export in absence of other ChAPs. (E) Fluorescence images of cells expressing Chs3‐GFP in WT, CHS6 ΔΔΔ, and GPD‐CHS6 ΔΔΔ strains. Arrows point to Chs3‐GFP signals at the bud neck. Scale bar, 5 μm. The inset represents a twofold magnification. (F) Quantification of phenotypes in (E); 100 small‐budded and 100 large‐budded cells were quantified per experiment. Average and SD of three independent biological experiments are shown. (G) Chs3 GFP GPD‐CHS6 ΔΔΔ strain was sensitive to calcofluor to a similar extent as the WT strain. Plates were incubated at 30°C for 2–3 days. A representative drop test of three independent biological experiments is shown.