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Several studies with different cell types showed that the TGN and Golgi stacks can contain plasma-membrane constituents and internalized materials (Stoorvogel et al. 1988; van Weert et al. 1997).
Low pH is critical for the degradation of internalized materials, the trafficking and fusion of endolysosomes and the formation of autophagy (Marshansky and Futai, 2008; Ravikumar et al., 2010; Williamson et al., 2010).
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Some of the internalized material is delivered to the degradative vacuole after passage through early and late endosomes.
Eukaryotic cell processing of internalized material classically involves transport from early to late endosomes, and then to lysosomes where degradation typically occurs.
Early endosomes bud from sorting endosomes and transport internalized material to lysosomes or recycling endosomes.
Many of these were complex with lipid whorls and internalized material, including mitochondria, peroxisomes and rough endoplasmic reticulum, in various stages of digestion.
Our observations of elevated endolysosome pH could help to explain alterations in the digestive capability of endolysosomes as evidenced by decreased specific activities of three different endolysosome enzymes, which results in increased accumulation of internalized material thus altering the structure and size of endolysosomes.
Statistical CCK-8 cell viability assay probably only reflected the cell that internalized hazardous materials rather than the overall destructiveness on cells.
TEM images of internalized carbonaceous material and SGSs within Hep3B liver cancer cells (A to F). Figure 4D,E,F is of the same cell.
lrrke03680 homozygous flies showed a similar, albeit weaker, accumulation of ubiquitylated proteins (supplementary material Fig. S4C versus S4B), which also colocalized with internalized dextran (supplementary material Fig. S4C″) and Hrs (supplementary material Fig. S4F″).
In order to characterize the endocytic pathway followed by PrPC, we developed an indirect immunofluorescence assay in cells grown in polarized conditions, which allowed us to distinguish between internalized and surface protein (Figure 1A, Materials and Methods).
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