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CARD19 interacts with MIB components and stabilizes mitochondrial cristae. (A) IP-MS results showing that CARD19 interacts with Myo18a, MIC60, SAMM50, and MIC19. (B) Representative immunoblot images showing that CARD19 interacts with MIC60, SAMM50, and MIC19 in microglia. (C) Representative immunoblot images showing that CARD19 overexpression partially rescued the binding between MIC19 and SAMM50 after OGD/R in microglia. (D) Representative immunoblot images showing that CARD19 knockdown of exacerbated the loss of binding between MIC19 and SAMM50 after OGD/R in microglia. All experiments were repeated at least three times, with each using a separate cell sample. CARD19: Caspase activation and recruitment domain 19; IP-MS: immunoprecipitation-mass spectrometry; MIB: mitochondrial intermembrane bridge; MIC19: mitochondrial inner membrane protein 19; MIC60: mitochondrial inner membrane protein 60; OGD/R: oxygen–glucose deprivation/reoxygenation; SAMM50: sorting and assembly machinery component 50.

Journal: Neural Regeneration Research

Article Title: Microglial CARD19 ameliorates post-stroke neuroinflammation by stabilizing mitochondrial cristae

doi: 10.4103/NRR.NRR-D-24-00923

Figure Lengend Snippet: CARD19 interacts with MIB components and stabilizes mitochondrial cristae. (A) IP-MS results showing that CARD19 interacts with Myo18a, MIC60, SAMM50, and MIC19. (B) Representative immunoblot images showing that CARD19 interacts with MIC60, SAMM50, and MIC19 in microglia. (C) Representative immunoblot images showing that CARD19 overexpression partially rescued the binding between MIC19 and SAMM50 after OGD/R in microglia. (D) Representative immunoblot images showing that CARD19 knockdown of exacerbated the loss of binding between MIC19 and SAMM50 after OGD/R in microglia. All experiments were repeated at least three times, with each using a separate cell sample. CARD19: Caspase activation and recruitment domain 19; IP-MS: immunoprecipitation-mass spectrometry; MIB: mitochondrial intermembrane bridge; MIC19: mitochondrial inner membrane protein 19; MIC60: mitochondrial inner membrane protein 60; OGD/R: oxygen–glucose deprivation/reoxygenation; SAMM50: sorting and assembly machinery component 50.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1.5 hours at room temperature, then incubated with primary antibodies against CARD19 (rabbit, 1:1000, Abclonal, Wuhan, China, Cat# A13196), GAPDH (rabbit, 1:2000, Bioworld Technology, St. Louis Park, MN, USA, Cat# MB66349), β-actin (rabbit, 1:5000, Bioworld Technology, Cat# AP0060), mitochondrial inner membrane protein 19 (MIC19) (rabbit, dilution: 1:1000, Abclonal, Cat# A8584), mitochondrial inner membrane protein 60 (MIC60) (rabbit, 1:1000, Abclonal, Cat# A2751), sorting and assembly machinery component 50 (SAMM50) (rabbit, 1:1000, Abclonal, Cat# A3401), and Flag (rabbit, dilution: 1:1000, Abclonal, Cat# AE063) overnight at 4°C.

Techniques: Protein-Protein interactions, Western Blot, Over Expression, Binding Assay, Knockdown, Activation Assay, Immunoprecipitation, Mass Spectrometry, Membrane