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Figure 2


Fig. 2. In vivo topology analysis of mAtg9. Plasmids YFP-HLA-A2-CFP and mRFPmAtg9 were transiently transfected into NRK cells. YFP on the N-terminus is in the ER lumen and CFP on its C-terminus is cytosolic. (A) Pre-treatment images. (B) Images captured after 2 minutes of permeabilization with digitonin. (C) Images captured 5 minutes after subsequent trypsin addition. At the concentration used, digitonin permeabilized the plasma membrane but not the ER. Lumenal YFP on the N-terminus of HLA-A2 was protected from the trypsin, but the C-terminal CFP and the mRFP on the N-terminus of mAtg9 were degraded by the trypsin and are therefore cytosolic. All images were captured using identical imaging parameters.





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