Journal: Experimental Animals

Article Title: Ectodysplasin-A2 receptor (EDA2R) knockdown alleviates myocardial ischemia/reperfusion injury through inhibiting the activation of the NF-κB signaling pathway

doi: 10.1538/expanim.24-0020

Figure Lengend Snippet: Dexmedetomidine (DEX) has a protective effect on hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury. (A) Cell -Counting Kit 8 (CCK-8) was used to measure cell viability. (B, C) Apoptosis and apoptotic rate were determined using flow cytometry. (D) GSE126104 showed that DEX decreased ectodysplasin-A2 receptor (EDA2R) expression in the left ventricle of rats. ***, P <0.001 vs. con. (E) The expression of EDA2R in cardiomyocytes was detected by RT-qPCR and western blotting. ** P <0.01; *** P <0.001; **** P <0.0001 vs. con. ## P <0.01; #### P <0.0001 vs. H/R. n=3.

Article Snippet: AC16 human cardiomyocytes were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Cell Counting, CCK-8 Assay, Flow Cytometry, Expressing, Quantitative RT-PCR, Western Blot

Journal: Experimental Animals

Article Title: Ectodysplasin-A2 receptor (EDA2R) knockdown alleviates myocardial ischemia/reperfusion injury through inhibiting the activation of the NF-κB signaling pathway

doi: 10.1538/expanim.24-0020

Figure Lengend Snippet: Ectodysplasin-A2 receptor (EDA2R) knockdown suppresses hypoxia/reoxygenation (H/R)-induced cardiomyocyte apoptosis. (A) The knockdown efficiency of EDA2R in cardiomyocytes was detected by RT-qPCR and western blotting. (B) Cell -Counting Kit 8 (CCK-8) was applied to detect cell viability. (C, D) Apoptosis was determined using flow cytometry, and apoptotic rate was calculated. *** P <0.001; **** P <0.0001 vs. con. ## P <0.01; ### P <0.001; #### P <0.0001 vs. H/R+shNC. n=3.

Article Snippet: AC16 human cardiomyocytes were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Cell Counting, CCK-8 Assay, Flow Cytometry

Journal: Experimental Animals

Article Title: Ectodysplasin-A2 receptor (EDA2R) knockdown alleviates myocardial ischemia/reperfusion injury through inhibiting the activation of the NF-κB signaling pathway

doi: 10.1538/expanim.24-0020

Figure Lengend Snippet: Ectodysplasin-A2 receptor (EDA2R) knockdown represses mitochondria-mediated apoptosis. (A) Mitochondrial morphology of cardiomyocytes was observed by transmission electron microscopy (the arrows represented mitochondrial morphology, scale bar=500 nm). (B) Mitochondrial membrane potential (MMP) was detected by JC-1 (Scale bar=100 µ m, Red: **** P <0.0001 vs. con. or hypoxia/reoxygenation (H/R)+shNC; Green: #### P <0.0001 vs. con. H/R+shNC). (C, D) The levels of cytochrome C in mitochondria and cytoplasm of cardiomyocytes were determined by western blotting. (E) The expressions of Bax and Bcl-2 were determined in cardiomyocytes by western blotting. (F, G) Caspase-3 and Caspase-9 activity in cardiomyocytes was detected. *** P <0.001; **** P <0.0001 vs. con. # P <0.05; ## P <0.01; ### P <0.001; #### P <0.0001vs. H/R+shNC. n=3.

Article Snippet: AC16 human cardiomyocytes were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Knockdown, Transmission Assay, Electron Microscopy, Membrane, Western Blot, Activity Assay

Journal: Experimental Animals

Article Title: Ectodysplasin-A2 receptor (EDA2R) knockdown alleviates myocardial ischemia/reperfusion injury through inhibiting the activation of the NF-κB signaling pathway

doi: 10.1538/expanim.24-0020

Figure Lengend Snippet: Ectodysplasin-A2 receptor (EDA2R) knockdown inhibits the activation of the NF-κB signaling pathway. (A) The levels of IκBα, p-IκBα (Ser32), NF-κB p65 and p-NF-κB p65 (Ser536) were detected in cardiomyocytes by western blotting. (B) The expression distribution of NF-κB p65 in cardiomyocytes was determined by immunofluorescence, the fluorescence intensity of p65 in the nucleus was quantified in three fields of three sections (the arrows represented the distribution of p65 in the nucleus, scale bar=50 µ m). (C) The transcriptional activity of NF-κB in cardiomyocytes was measured by the electrophoretic mobility shift assay (EMSA). **** P <0.0001 vs. con. #### P <0.0001. n=3.

Article Snippet: AC16 human cardiomyocytes were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Knockdown, Activation Assay, Western Blot, Expressing, Immunofluorescence, Fluorescence, Activity Assay, Electrophoretic Mobility Shift Assay