Figure 8. ATG9 compartments that... | Rockefeller University Press

$ATG9 compartments that co-localize with LC3 foci are lysosomes. (A) Micrographs showing the cellular distribution of transiently expressed RFP-ATG9A, endogenous Halo-ATG9A, and lysosomes, marked with mNeon-LAMP1. Experiments were performed immediately (upper panel) and 24 h (bottom panel) after labeling Halo-ATG9A. Scale bar = 10 μm. (B–E) Quantification of Halo-ATG9A and RFP-ATG9A tracks colocalized with mNeon-LAMP1. The number of cells (n) is indicated in each graph in the figure panels. Top schemes depict which signals were used to calculate the fractions in the corresponding graph. The marker represents mean ± 95% confidence interval. Letters indicate a statistically homogeneous group established by ANOVA (P < 0.05) followed by Bonferroni post-hoc test. (F) Micrographs showing the cellular distribution of endogenous edited SNAP-LC3 and Halo-ATG9A, and lysosomes, marked with LysoTracker. Experiments were performed immediately (upper panel) and 24 h (bottom panel) after Halo-ATG9A labeling. Scale bar = 10 µm. (G) Kymographs showing colocalization of SNAP-LC3/Halo-ATG9A immediately and 24 h after Halo-ATG9A labeling. (H) Number of co-diffusing Halo-ATG9A and SNAP-LC3 foci immediately and 24 h after Halo-ATG9A labeling. The marker represents mean ± 95% confidence interval (*P < 0.05). The number of data points (n) is indicated in each graph in the figure panels. (I) Fraction of colocalized SNAP-LC3 and Halo-ATG9A foci that also colocalized with lysosomes marked with LysoTracker. The marker represents mean ± 95% confidence interval.$

Figure 8.

ATG9 compartments that co-localize with LC3 foci are lysosomes. (A) Micrographs showing the cellular distribution of transiently expressed RFP-ATG9A, endogenous Halo-ATG9A, and lysosomes, marked with mNeon-LAMP1. Experiments were performed immediately (upper panel) and 24 h (bottom panel) after labeling Halo-ATG9A. Scale bar = 10 μm. (B–E) Quantification of Halo-ATG9A and RFP-ATG9A tracks colocalized with mNeon-LAMP1. The number of cells (n) is indicated in each graph in the figure panels. Top schemes depict which signals were used to calculate the fractions in the corresponding graph. The marker represents mean ± 95% confidence interval. Letters indicate a statistically homogeneous group established by ANOVA (P < 0.05) followed by Bonferroni post-hoc test. (F) Micrographs showing the cellular distribution of endogenous edited SNAP-LC3 and Halo-ATG9A, and lysosomes, marked with LysoTracker. Experiments were performed immediately (upper panel) and 24 h (bottom panel) after Halo-ATG9A labeling. Scale bar = 10 µm. (G) Kymographs showing colocalization of SNAP-LC3/Halo-ATG9A immediately and 24 h after Halo-ATG9A labeling. (H) Number of co-diffusing Halo-ATG9A and SNAP-LC3 foci immediately and 24 h after Halo-ATG9A labeling. The marker represents mean ± 95% confidence interval (*P < 0.05). The number of data points (n) is indicated in each graph in the figure panels. (I) Fraction of colocalized SNAP-LC3 and Halo-ATG9A foci that also colocalized with lysosomes marked with LysoTracker. The marker represents mean ± 95% confidence interval.

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