May gr nwald and giemsa stain solution

Bovine aortic endothelial cells were purchased from Cell Applications (San Diego, CA, USA). The following materials were purchased from the respective vendors: Dulbecco's modified Eagle's medium (DMEM) and calcium-and magnesium-free phosphate-buffered saline (CMF-PBS; Nissui Pharmaceutical, Tokyo, Japan); fetal bovine serum and a High-Capacity cDNA Reverse Transcription kit (Thermo Fisher Scientific, Waltham, MA, USA); QIAzol lysis reagent (QIAGEN, Valencia, CA, USA); GeneAce SYBR qPCR Mixα (Nippon Gene, Tokyo, Japan); mouse monoclonal anti-MT-1/2 antibody (E9; Dako, Glostrup, Denmark); Mouse monoclonal anti-β-actin antibody (Wako Pure Chemical Industries, Osaka, Japan); horseradish peroxidase-conjugated antimouse IgG antibody (#7076; Cell Signaling, Beverly, MA, USA); May-Grünwald and Giemsa stain solution (Merck KGaA, Darmstadt, Germany); and cadmium chloride, manganese chloride tetrahydrate, and other reagents (Nacalai Tesque, Kyoto, Japan).

Bovine aortic endothelial cells and smooth muscle cells were purchased from Cell Applications (San Diego, CA, USA). Epithelial LLC-PK 1 cells derived from porcine kidney and fibroblastic IMR-90 cells derived from human fetal lung were purchased from DS Pharma Biomedical (Osaka, Japan). The zinc complexes used in this study were supplied by Takehiro Nakamura, a research associate at Kindai University, Osaka, Japan. The following materials were purchased from the respective vendors: Dulbecco's modified Eagle's medium (DMEM) and calcium-and magnesium free-phosphate buffered saline (CMF-PBS) from Nissui Pharmaceutical (Tokyo, Japan); fetal bovine serum from Thermo Fisher Scientific (Waltham, MA, USA); and tissue culture plates and dishes from AGC Techno Glass (Shizuoka, Japan). Other materials include CytoTox 96 Non-Radioactive Cytotoxicity Assay, a lactate dehydrogenase kit, from Promega (Madison, WI, USA); May-Grünwald and Giemsa stain solution from Merck KGaA (Darmstadt, Germany); AlamarBlue, a cell viability assay reagent from Bio-Rad Laboratories (Hercules, CA, USA); 3,5-diaminobenzoic acid from Tokyo Chemical Industry (Tokyo, Japan); nitric oxide and other reagents from Nacalai Tesque (Kyoto, Japan).