A HaloTag-based platform for quantitative analysis of autophagy in human cells. (A) Model showing autophagy factors and the complexes they form from phagophore initiation, toward phagophore expansion and autophagosome closure. Proteins tagged in this study are indicated in pink. (B) Western blots of autophagy proteins showing size shift of the tagged protein and exclusive expression of the tagged protein in comparison to the parental U2OS cell line. Three concentrations (100, 50, and 25% of initial lysis volume) were loaded on the gel. (C) Fluorescence gel showing gene tagging; two distinct monoclonal lines (C1 and C2) were selected for each edited gene. Cell lines were labeled with saturating amounts of HaloTag ligand JF646 (250 nM, 30 min). Source data are available for this figure: SourceData F1.
Figure 1.

A HaloTag-based platform for quantitative analysis of autophagy in human cells. (A) Model showing autophagy factors and the complexes they form from phagophore initiation, toward phagophore expansion and autophagosome closure. Proteins tagged in this study are indicated in pink. (B) Western blots of autophagy proteins showing size shift of the tagged protein and exclusive expression of the tagged protein in comparison to the parental U2OS cell line. Three concentrations (100, 50, and 25% of initial lysis volume) were loaded on the gel. (C) Fluorescence gel showing gene tagging; two distinct monoclonal lines (C1 and C2) were selected for each edited gene. Cell lines were labeled with saturating amounts of HaloTag ligand JF646 (250 nM, 30 min). Source data are available for this figure: SourceData F1.

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