Figure 3. Pex10, Pex11, and Pmp47 do not... | Rockefeller University Press

Figure 3.

$Figure 3. Pex10, Pex11, and Pmp47 do not colocalize with Pex14. (A) FM images of pex3 atg1 cells grown for 16 h on MM-M/G. Cells produced Pex14-mCherry and C-terminal mGFP fusions of the indicated proteins. (B) WB analysis of WT (1) and pex3 atg1 cells (2), grown for 6 or 16 h on MM-M/G. (C) FM images showing mGFP-fluorescence in pex3 atg1 cells producing Pex14-mCherry (control) or Pex14-mCherry together with the indicated mGFP fusion protein. Cells were grown for 6 h on MM-M/G. (D and E) iEM of pex3 atg1 cells using α-Pex5 (D) or α-alcohol oxidase antibodies (E). (F) Cell fractionation of the two indicated strains (seperated by the horizontal broken line). Postnuclear supernatants (PNS) were subjected to differential centrifugation resulting in a 30,000 g organelle pellet (P) and supernatant fraction (S). (G) Flotation analysis of the organelle pellet showing the distribution of the indicated proteins in the top (1) to bottom (10) fractions. (H) Colocalization of Pex8-GFP and Pex14-mCherry in pex10 cells. Bars: (A) 2.5 µm; (C) 5 µm; (D and E) 100 nm. AOX, cytosolic alcohol oxidase crystalloid; N, nucleus; V, vacuole.$

Pex10, Pex11, and Pmp47 do not colocalize with Pex14. (A) FM images of pex3 atg1 cells grown for 16 h on MM-M/G. Cells produced Pex14-mCherry and C-terminal mGFP fusions of the indicated proteins. (B) WB analysis of WT (1) and pex3 atg1 cells (2), grown for 6 or 16 h on MM-M/G. (C) FM images showing mGFP-fluorescence in pex3 atg1 cells producing Pex14-mCherry (control) or Pex14-mCherry together with the indicated mGFP fusion protein. Cells were grown for 6 h on MM-M/G. (D and E) iEM of pex3 atg1 cells using α-Pex5 (D) or α-alcohol oxidase antibodies (E). (F) Cell fractionation of the two indicated strains (seperated by the horizontal broken line). Postnuclear supernatants (PNS) were subjected to differential centrifugation resulting in a 30,000 g organelle pellet (P) and supernatant fraction (S). (G) Flotation analysis of the organelle pellet showing the distribution of the indicated proteins in the top (1) to bottom (10) fractions. (H) Colocalization of Pex8-GFP and Pex14-mCherry in pex10 cells. Bars: (A) 2.5 µm; (C) 5 µm; (D and E) 100 nm. AOX, cytosolic alcohol oxidase crystalloid; N, nucleus; V, vacuole.