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. 2019 Aug 21;20(10):e48014. doi: 10.15252/embr.201948014

Figure 6. UBE2QL1 is essential for efficient lysophagy and cell survival after lysosomal damage.

Figure 6

  1. Gal3 puncta assay for damaged lysosomes. HeLa cells were depleted of UBE2QL1 with two siRNAs for 60 h and treated with LLOMe or EtOH alone (untreated) for 1 h. Cells were fixed at indicated times after washout, stained with DAPI and Gal3 and LAMP1 antibodies, and processed for confocal microscopy. Note increased number of Gal3 puncta in untreated UBE2QL1‐depleted cells and their persistence 10 h after LLOMe‐induced damage. Scale bars: 10 μm, 2 μm for inlays. Arrows indicate Gal3 and LAMP1 colocalizing vesicles.
  2. Automated quantification of (A). The percentage of cells with more than three Gal3 puncta is shown. Graph represents data from three independent experiments with ≥ 50 cells per condition (mean ± SD). *P < 0.05; **P < 0.01 (one‐way ANOVA with Dunnett's multiple comparison test).
  3. Survival assay. Control or UBE2QL1‐depleted HeLa cells (48 h) were treated with increasing concentrations of LLOMe as indicated. Cell viability was measured with the MTS assay. Graph represents data from one experiment including three replicates (mean ± SD). *P < 0.05; ****P < 0.0001 (one‐way ANOVA with Bonferroni's multiple comparison test).