Journal: The Journal of Cell Biology

Article Title: Inhibitors of Copi and Copii Do Not Block PEX3 -Mediated Peroxisome Synthesis

doi:

Figure Lengend Snippet: Alignment of human, CHO, and yeast homologues of SAR1. Identical residues are shaded, and the asterisk identifies threonine 39. The sequence of HsSAR1 is available from GenBank/EMBL/DDBJ under accession number AF261717.

Article Snippet: Clones encoding the human SAR1 were obtained from Genome Systems Inc., and were sequenced in their entirety.

Techniques: Sequencing

Journal: The Journal of Cell Biology

Article Title: Inhibitors of Copi and Copii Do Not Block PEX3 -Mediated Peroxisome Synthesis

doi:

Figure Lengend Snippet: PEX3 targeting to peroxisomes is independent of COPII-mediated ER export. Normal human fibroblasts were cotransfected with either pcDNA3- GALT3xHA and pcDNA3 (A) or pcDNA3- GALT3xHA and pcDNA3- SAR1/T39N/vsvg (B and C) and processed 1 d later for indirect immunofluorescence with antibodies to the HA epitope (A and B) and the vsvg epitope (C). Note the accumulation of GALT in the ER in cells expressing the SAR1 mutant. Normal human fibroblasts were also transfected with pcDNA3- SAR1/T39N/vsvg , incubated for 1 d, and then microinjected with pcDNA3- PEX3myc (D and E). 1 h later, the cells were processed for double indirect immunofluorescence with antibodies to the myc epitope (D) and the vsvg epitope (E). Note that PEX3 is still targeted to peroxisomes, even in cells expressing high level of the SAR1 mutant. Bar, 10 μm.

Article Snippet: Clones encoding the human SAR1 were obtained from Genome Systems Inc., and were sequenced in their entirety.

Techniques: Immunofluorescence, Expressing, Mutagenesis, Transfection, Incubation

Journal: The Journal of Cell Biology

Article Title: Inhibitors of Copi and Copii Do Not Block PEX3 -Mediated Peroxisome Synthesis

doi:

Figure Lengend Snippet: PEX3 -mediated peroxisome synthesis is independent of COPII. PDB400 cells were cotransfected with either pcDNA3- GALT3xHA and pcDNA3 (A) or pcDNA3- GALT3xHA and pcDNA3- SAR1/T39N/vsvg (B and C), and were then processed 2 d later for indirect immunofluorescence with antibodies to the HA epitope (A and B) and the vsvg epitope (C). PBD400 cells were also cotransfected with pcDNA3- SAR1/T39N/vsvg and pcDNA3- PEX3 (D and E) and processed 2 d later for double indirect immunofluorescence with antibodies to PMP70 (D) and the vsvg epitope (E). Note that in PBD400 cells expressing the SAR1 mutant, GALT accumulates in the ER, yet PEX3 is still able to rescue peroxisome synthesis. Bar, 10 μm.

Article Snippet: Clones encoding the human SAR1 were obtained from Genome Systems Inc., and were sequenced in their entirety.

Techniques: Immunofluorescence, Expressing, Mutagenesis