Figure 2. UBE2QL1 depletion affects ubiquitination of damaged lysosomes.

• A
  HeLa cells were transfected with different UBE2QL1 siRNAs for 60 h, and LLOMe or control treated (untreated) for 3 h. Cells were fixed and immunostained for LAMP1 along with antibodies specific for K63 or K48‐linked ubiquitin chains as indicated and imaged with a laser scanning confocal microscope. Note the strong reduction of damage‐induced K48 ubiquitination and decreased intensity of the K63 signal. Scale bar: 20 μm.
• B, C
  Automated image quantification of (A). Shown are percentages of K48 or K63‐positive LAMP1 vesicles per cell. Graphs represent data from three independent experiments with ≥ 70 cells per condition (mean ± SD). **P < 0.01; ***P < 0.001; ****P < 0.0001 (one‐way ANOVA with Bonferroni's multiple comparison test).