Phosphodiesterase 2A2 regulates mitochondria clearance through Parkin-dependent mitophagy

摘要

a Volcano plot of the PDE2A2 interactome. Filled circles indicate proteins significantly pulled-down with PDE2A2. Highlighted in orange are proteins annotated to the GOCC category “Mitochondrial membrane” and in blue are proteins annotated to the category ‘Mitochondrial matrix’. PDE2A2, MICOS components and MICOS-interacting proteins are shown in red. b Confocal images of Sh-Sy5y cells expressing PDE2A2-GFP. Cells were fixed and immunostained with GFP- (red), MIC60- (turquoise) specific antibodies. Images are representative of two independent experiments. Scale bar: 10 μm. c Detection of endogenous MIC60 after PDE2A2-RFP or mRFP immunoprecipitation from lysates obtained from HEK293T cells transiently expressing either PDE2A2-RFP or mRFP. d Detection of endogenous MIC19 after PDE2A2-RFP or mRFP immunoprecipitation from lysates obtained from HEK293T cells transiently expressing either PDE2A2-RFP or mRFP. e Detection of endogenous SAMM50 after PDE2A2-GFP or GFP immunoprecipitation from lysates obtained from HEK293T cells transiently expressing either PDE2A2-GFP or GFP. f Detection of endogenous PDE2A after immunoprecipitation of endogenous MIC60 from rat brain cell lysates. g Detection of endogenous MIC60 after immunoprecipitation of each PDE2A isoform from lysates obtained from HEK293T cells transiently expressing PDE2A1-GFP, PDE2A2-GFP, PDE2A3-GFP or PDE2A4-GFP. PDE2A4, a variant whose sequence has been deposited (NCBI Reference Sequence: {"type":"entrez-nucleotide","attrs":{"text":"NM_001146209.2","term_id":"344925849","term_text":"NM_001146209.2"}}NM_001146209.2) but for which no further characterization is available, was used here as a control. All blots are representative of three independent experiments except from data shown in (f), which is representative of two independent experiments. WCL whole-cell lysate.