Apc conjugated anti pdgfr α

Cardiac fibroblasts (PDGFR α⁺) and endothelial cells (CD31 ⁺) were isolated from wild-type C57Bl/6J mouse hearts and sorted using APC-conjugated anti-PDGFR α⁺ (eBioscience 17-1401-81) and PE-CY7-conjugated anti-CD31 (eBioscience 25-0311-82) antibodies as described previously [37 (link)]. Cardiomyocytes were isolated from wild-type C57Bl/6J mice as described previously [53 (link)].

Six male C57BL/6 mice each from the control and cinnamtannin B-1-treated groups were injected with 0.4 mL phosphate-buffered saline (PBS) or cinnamtannin B-1 solution (2.0 mg/kg via the vena caudalis), respectively. The experiments were performed five times. After 1 h, approximately 1 mL samples of peripheral blood were collected, and nucleated cells were separated using Ficoll gradient centrifugation (final density, 1.077 g/m). To detect the endogenous MSCs with the lineage-negative, PDGFRα+, and Sca-1+ (Lin-/PDGFRα+/Sca-1+) cells were stained with allophycocyanin (APC)-conjugated anti-PDGFRα (eBioscience, San Diego, CA, USA), PE-conjugated anti-Sca-1 (eBioscience), or V450 Mouse Lineage Antibody Cocktail (BD Biosciences, San Jose, USA) diluted in PBS and fixed with 1% paraformaldehyde. Flow cytometry analysis was performed using a FACSCanto II flow cytometer (BD Biosciences) using FACSDiva software. The percentage of peripheral blood mononuclear fraction cells that targeted the Lin-/PDGFRα+/Sca-1+ in the control and cinnamtannin B-1 groups was compared.