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. 2021 Feb 10;22(3):e49097. doi: 10.15252/embr.201949097

Figure EV4. MITOL translocates to the ER in an FKBP38‐dependent manner, related to Fig 3 .

Figure EV4

  • A
    MITOL translocates from the mitochondria to the ER in the late phase of mitophagy with FKBP38. HeLa cells stably expressing HA‐Parkin were transfected with the indicated vectors and treated with DMSO or CCCP (10 μM) for 48 h. Cells were fixed, permeabilized, and subjected to immunofluorescence analysis with indicated antibodies. Colocalization was quantified by Manders’s coefficient. Means ± SEM of more than 10 cells obtained from three independent experiments. For statistical analysis, Student’s t‐tests were performed, ****P < 0.0001. Scale bar represents 10 μm. Higher magnification images of the boxed regions are shown in the small panel.
  • B
    Interaction of endogenous MITOL and FKBP38. HeLa cells stably expressing HA‐Parkin were treated with DMSO or CCCP (10 μM) for indicated times and subjected to an IP‐IB assay with the indicated antibodies.
  • C, D
    MITOL translocation depends on FKBP38. HeLa cells stably expressing HA‐Parkin were transfected with FKBP38 siRNA or control siRNA and treated with DMSO or CCCP (10 μM) for 30 h. Lysates of cells were fractioned into whole‐cell, cytosolic, ER, and mitochondrial fractions and then subjected to an IB assay with the indicated antibodies (C). MITOL KO HeLa cells stably expressing HA‐Parkin were transfected with MITOL‐myc and treated with DMSO or CCCP (10 μM) for 48 h. Cells were immunostained with indicated antibodies (D). Scale bar, 10 μm.