Fig. 2: KANSL1 deficiency impairs autophagosome-lysosome fusion. | Nature Communications

Fig. 2: KANSL1 deficiency impairs autophagosome-lysosome fusion.

From: Kansl1 haploinsufficiency impairs autophagosome-lysosome fusion and links autophagic dysfunction with Koolen-de Vries syndrome in mice

Fig. 2

a Western blot analysis of HeLa cells transfected with siRNAs with the indicated antibodies. Cells were treated with or without 100 nM bafilomycin A1 for 12 h. The arrow indicated the KANSL1 protein band. b Quantitation of protein signal intensities from immunoblots in (a) showing the ratio of LC3B-II to LC3B-I (n = 3 independent experiments). c Representative images of RFP-GFP-LC3B lentivirus-infected HeLa cells transfected with the indicated siRNAs following EBSS (6 h) treatment. Nuclei are labeled with DAPI (blue). Scale bar, 10 μm. d, e Quantification of the number of autolysosome (d) or autophagosome (e) per cell in (c). n = 82, 83, 77 cells. f Representative confocal images of colocalization of GFP-LC3B puncta and acid lysosome (red, LysoTracker) in HeLa cells transfected with the indicated siRNAs after EBSS treatment for 6 h. g, h Quantification of the colocalization by calculating the percentage of LC3B+ Lyso+ puncta to total LC3B puncta (g) and Pearson’s correlation coefficient (h) in (f). n = 61, 58, 42 cells. Scale bar, 10 μm and 1 μm in “Zoom”. Source data are provided as a Source data file. All data are means ± SEM. b One-way ANOVA with Tukey’s multiple hoc test; d, e, g, h by one-way ANOVA with Dunnett’s multiple hoc test.

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