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Image Search Results
Journal: Frontiers in Cell and Developmental Biology
Article Title: Applying Sodium Carbonate Extraction Mass Spectrometry to Investigate Defects in the Mitochondrial Respiratory Chain
doi: 10.3389/fcell.2022.786268
Figure Lengend Snippet: Loss of UQCR10 leads to destabilization and accumulation of a soluble core-module. (A) Mitochondria isolated from indicated cell lines were solubilized in 1% digitonin and analyzed by BN-PAGE and immunoblotting with the indicated antibodies. Ŧ, † and ‡ represent sub-assemblies discussed in text. *non-specific complex. (B) Oxygen consumption rates of HEK293T, UQCRC1 KO and UQCR10 KO cell lines. Oligo, oligomycin; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; Ant A, antimycin A; Rot, rotenone. (C) Mitochondria isolated from indicated cell lines were analyzed using BN-PAGE and the activity of complex I and IV determined through in-gel assays. (D) Volcano plot depicting relative abundances of mitochondrial proteins in UQCR10 KO compared to HEK293T. (E) Topographical heatmap showing changes in abundance of CIII and respirasome subunits in UQCR10 KO using the CIII (PDB 5XTE) and respirasome (PDB 5XTH) structures. UQCR10 shown in yellow. (F) Volcano plots depicting relative abundance of proteins in pH 9.5 carbonate extraction pellet and supernatant fractions in UQCR10 KO compared to HEK293T mitochondria following normalization against the total fraction. Data prior to normalization and pH 11.5 data can be found in . (G) Principal component analysis showing overall membrane association profiles in UQCR10 KO relative to HEK293T mitochondria. Complex I N-module and CIII subunits are labeled, with their position in control data shown as empty squares and position in knockout data as filled squares. Clusters defined in are colored in grey.
Article Snippet: Western transfer of
Techniques: Isolation, Western Blot, Activity Assay, Labeling, Knock-Out