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. 2015 Jul 2;34(17):2272–2290. doi: 10.15252/embj.201590992

Figure 6.

Figure 6

UVRAG phosphorylation alters lysosomal morphology

  1. U2OS cells or those stably expressing wild-type (WT) or S550A+S571A (dblA) GFP-UVRAG were transfected with 100 nM control or UVRAG siRNA 40 h prior to treatment. Cells were grown in complete medium prior to fixation and staining for LAMP1. Scale bar, 10 μm.
  2. Western blot of endogenous UVRAG depletion and rescue with GFP-UVRAG as described in (A).
  3. Quantitation of LAMP1 punctate structures and size from (A), mean ± SEM for n = 3 independent experiments, significance was determined by one-way ANOVA and Dunnett’s multiple comparison test to the siCTRL, ***P < 0.001 and n.s = not significant.
  4. U2OS cells or those stably expressing wild-type (WT) or S550A+S571A (dblA) GFP-UVRAG were transfected with 100 nM control or UVRAG siRNA 40 h prior to treatment. Cells were grown in complete medium prior to fixation and staining for LAMP1 and PX domain. Scale bar, 10 μm. Arrowheads indicate co-localising punctate structures.
  5. Quantitation of LAMP1 co-localisation with PX domain as in (D) ± SEM from n = 3 independent experiments. Significance was determined by one-way ANOVA and Dunnett’s multiple comparison test to the siCTRL, **P < 0.01.

Source data are available online for this figure.