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. 2020 Jul 9;39(18):e104494. doi: 10.15252/embj.2020104494

Figure 5. Damaged endolysosomes are targeted to lysophagy in the absence of LRRK2 and Rab8A signalling.

Figure 5

  • A, B
    WT or LRRK2 KO RAW264.7 macrophages, RAW264.7 macrophages pre‐treated with 1 μM GSK2578215A (GSK inh) and WT or Rab8A KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min. (A) LC3B and (B) ubiquitin K63‐positive vesicle numbers were analysed by immunofluorescence and high‐content imaging. Scale bar = 20 μm. Right panels show quantification of number of positive vesicles per cell. Data show the mean ± SEM of three to four biological replicates. ns = non‐significant, *P ≤ 0.05, **P ≤ 0.01 by one‐way ANOVA followed by Sidak's multiple comparisons test.
  • C
    Representative images of LC3B/LAMP‐1 double‐positive endolysosomes in control and LLOMe‐treated macrophages. Scale bar = 10 μm.
  • D
    Analysis of the % of LC3B/LAMP‐1 double‐positive endolysosomes in LLOMe‐treated macrophages. Data show mean ± SEM of three independent biological experiments. *P ≤ 0.05, ***P ≤ 0.001 by Student's t‐test.