Journal: The Journal of Experimental Medicine

Article Title: The tight junction protein CAR regulates cardiac conduction and cell–cell communication

doi: 10.1084/jem.20080897

Figure Lengend Snippet: Altered expression of the cell–cell contact proteins in CAR KO hearts. (A) Expression of the CAR and its adaptor protein ZO-1 were reduced significantly from weeks 2 and 4, respectively, after induction of the KO. (B) The composition of the GAP junction was altered as a result of differential expression of Cx37 (transiently reduced) and Cx40 (transiently increased, albeit not significantly). Expression of Cx43 was unchanged. (C) Cx45 was the only connexin with RNA levels significantly altered late in the progression of the phenotype (reduced from >8 wk). The adherens junction protein N-cadherin was unchanged. All expression data were normalized to GAPDH and levels at week 0 (before induction) were set to 100%. Fold change is shown on the y-axis. n = 3 per group. Error bars show SD. (D) Cardiac expression of Cx40, Cx43, and Cx45 was confirmed on the protein level with down-regulation of Cx43 and Cx45, but not Cx40, in animals at >8 wk after induction of the phenotype. GAPDH was used as a loading control. After normalization to GAPDH, Cx45 levels were reduced by 45% (P = 0.012), Cx 43 levels by 43% (P = 0.015), and Cx40 levels were not changed significantly ( n = 4). *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.

Article Snippet: Primary antibodies used against CAR (rabbit polyclonal; Santa Cruz Biotechnology, Inc.), α-actin (rabbit polyclonal; Sigma-Aldrich), Cx40 (rabbit polyclonal; Santa Cruz Biotechnology, Inc.), Cx45 (mouse monoclonal; Invitrogen), and GAPDH (mouse monoclonal; Affinity BioReagents) were used according to the manufacturer's instructions and detected with HRP-conjugated secondary antibodies and chemiluminescence staining using ECL (Supersignal West Pico Chemiluminescent Substrate; Thermo Fisher Scientific).

Techniques: Expressing, Quantitative Proteomics, Control

Journal: The Journal of Experimental Medicine

Article Title: The tight junction protein CAR regulates cardiac conduction and cell–cell communication

doi: 10.1084/jem.20080897

Figure Lengend Snippet: Conduction phenotypes with prolonged PR interval

Article Snippet: Primary antibodies used against CAR (rabbit polyclonal; Santa Cruz Biotechnology, Inc.), α-actin (rabbit polyclonal; Sigma-Aldrich), Cx40 (rabbit polyclonal; Santa Cruz Biotechnology, Inc.), Cx45 (mouse monoclonal; Invitrogen), and GAPDH (mouse monoclonal; Affinity BioReagents) were used according to the manufacturer's instructions and detected with HRP-conjugated secondary antibodies and chemiluminescence staining using ECL (Supersignal West Pico Chemiluminescent Substrate; Thermo Fisher Scientific).

Techniques: Blocking Assay, Isolation

Journal: iScience

Article Title: CCRR regulate MYZAP-PKP2-Nav1.5 signaling pathway in atrial fibrillation following myocardial infarction

doi: 10.1016/j.isci.2024.111102

Figure Lengend Snippet:

Article Snippet: The antibodies used were PKP2 (1:500, abs136897, Absin, China), TLR2 (1:2000, DF7521, Affinity, America), TLR4 (1:2000, 66350-1, Proteintech, America), MYZAP (1:2000, PA5-21116, Invitrogen, America), CX40 (1:1000, DF13633, Affinity, America), CX43 (1:1000, 26980-1, Proteintech, America), Nav1.5 (1:200, ASC-005, Alomone labs, Israel), Cav1.2 (1:200, ACC-003, Alomone labs, Israel), Kv4.2 (1:200, APC-023, Alomone labs, Israel), β-actin(1:3000, A00702, GenScript, America), β-tubulin(1:2000, WL01931, Wanleibio, China) and GAPDH (1:2000, BA002, Bio-Platform, China).

Techniques: Virus, Recombinant, Bicinchoninic Acid Protein Assay, CCK-8 Assay, Immunoprecipitation, SYBR Green Assay, Software, Sequencing