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

Figure EV4. Characterisation of Rab GTPase behaviour after sterile damage.

Figure EV4

  1. RAW264.7 macrophages were electroporated with EGFP‐Rab3A, EGFP‐Rab8A, EGFP‐Rab10 and EGFP‐Rab35. Cells were treated with 1 mM of LLOMe for 30 min, and Rab recruitment to LAMP1 + compartments was monitored by high‐content immunofluorescence imaging. Scale bar = 10 μm.
  2. WT and LRRK2 KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min. Cells were separated into cytosolic (C) and membrane (M) fractions and analysed for Rab8A pT72, Rab8A and Rab10 by Western blot.
  3. WT and Rab8A KO RAW264.7 macrophages were treated with 1 mM LLOMe for 30 min, and Rab8A and Rab8A pT72 levels were analysed by Western blot.
  4. RAW264.7 macrophages were electroporated with EGFP‐Rab8A‐WT, EGFP‐Rab8A‐Q67L, EGFP‐Rab8A‐T22N and EGFP‐Rab8A‐T72A. Cells were treated with 1 mM of LLOMe for 30 min, and CHMP4B recruitment was assessed by confocal microscopy. Scale bar = 5 μm.
  5. CHMP4B integrated fluorescence density was analysed per cell. Data show values from single cells and mean.

Source data are available online for this figure.