LC3 subfamily in cardiolipin-mediated mitophagy: a comparison of the LC3A, LC3B and LC3C homologs [Dataset]
9 pages. -- Figure S1. LC3 binding to liposomes is enhanced by membrane curvature and, in a dose-dependent manner, by CL. Interaction of LC3 proteins with vesicles of different radii and/or CL contents was measured by a vesicle flotation assay. -- Figure S2. Increasing ionic strength of the medium d...
| Autores: | , , , , , |
|---|---|
| Tipo de recurso: | conjunto de datos |
| Fecha de publicación: | 2022 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/331563 |
| Acceso en línea: | http://hdl.handle.net/10261/331563 |
| Access Level: | acceso abierto |
| Palabra clave: | Atg8 Autophagosome Autophagy cargo recognition LC3/GABARAP-protein family Lipid oxidation Lipid-protein interaction Membrane curvature Mitochondria Negatively charged phospholipids |
| Sumario: | 9 pages. -- Figure S1. LC3 binding to liposomes is enhanced by membrane curvature and, in a dose-dependent manner, by CL. Interaction of LC3 proteins with vesicles of different radii and/or CL contents was measured by a vesicle flotation assay. -- Figure S2. Increasing ionic strength of the medium decreases the binding of LC3 to CL. Effect of the increased ionic strength on the interaction of LC3 proteins with CL-containing membranes, measured by a vesicle flotation assay. -- Figure S3. Changing a specific LC3C N-terminal residue has no effect on the protein binding to CL. -- Figure S4. Autophagy quantification with native GFP-LC3/GABARAP proteins and with non-conjugatable mutants confirms that most puncta in cells are autophagic vesicles and not aggregates. -- Figure S5. LC3A puncta and their colocalization with mitochondria increase with rotenone and CCCP treatment. Cells were co-transfected with DsRed2-Mito7 (DsRed) and with GFP-tagged WT or mutant LC3A. Vehicle (Veh) controls were treated with DMSO. -- Figure S6. The LC3B-AK double mutation that increases LC3B binding to CL in vitro does not have a comparable effect in cells. Cells were transfected with GFP-tagged WT or mutant LC3 protein. Mitochondria were labeled using MitoTracker Red, prior to the treatments. Vehicle (Veh) controls were treated with DMSO. -- Figure S7. mRNA and protein expression of LC3A, LC3B and LC3C in SH-SY5Y cells. -- Table S1. Primers used to perform site-directed mutagenesis. -- Table S2. Primers used to perform RT-qPCR. -- Table S3. siRNA synthesized by IDT to silence LC3A and LC3B proteins. |
|---|