Rat aortic smooth muscle cell lines (A7r5, ATCC) were cultured with low-glucose (1.0 g/L) Dulbecco’s Modified Eagle Medium (Wako) containing 10% (v/v) heat-inactivated fetal bovine serum (SAFC Biosciences) and 1% penicillin-streptomycin (Wako) in a humidified 5% CO2 incubator at 37°C. Expression plasmids encoding mClover2-tagged β-actin were constructed by inserting a human β-actin gene, which was digested with XhoI and BamHI restriction enzymes from the EYFP-actin vector (#6902–1, Clontech) into the mClover2-C1 vector (Addgene plasmid #54577, a gift from Michael Davidson) [16 (link)]. The plasmids were transfected to cells using Lipofectamin LTX with Plus Reagent (Thermo Fischer Science) according to the manufacturer’s instruction. Myosin II ATPase inhibitor (-)-blebbistatin (Wako) was used at 10 μM concentration, and actin polymerization inhibitor Latrunculin A (Wako) was used at 10 nM concentration. An equivalent amount of dimethyl sulfoxide (DMSO) was administered to cells as vehicle control.
Myosin 2 atpase inhibitor blebbistatin
Manufactured by Fujifilm
Myosin II ATPase inhibitor (-)-blebbistatin is a chemical compound used in laboratory research. It functions by selectively inhibiting the ATPase activity of myosin II, a motor protein involved in cellular processes such as muscle contraction and cell division.
Automatically generated - may contain errors
2 protocols using myosin 2 atpase inhibitor blebbistatin
1
Visualizing Actin Dynamics in Smooth Muscle Cells
2
Rat Aortic Smooth Muscle Cell Imaging
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Rat aortic smooth muscle cell lines (A75r, ATCC) were cultured with low-glucose (1.0 g/L)
Dulbecco's Modified Eagle Medium (Wako) containing 10% (v/v) heat-inactivated fetal bovine serum (SAFC Biosciences) and 1% penicillin-streptomycin (Wako) in a humidified 5% CO2 incubator at 37℃. Expression plasmids encoding mClover2-tagged β-actin were constructed by inserting a human β-actin gene, which was digested with XhoI and BamHI restriction enzymes from the EYFP-actin vector (#6902-1, Clontech) into the mClover2-C1 vector (Addgene plasmid #54577, a gift from Michael Davidson). The plasmids were transfected to cells using Lipofectamin LTX with Plus Reagent (Thermo Fischer Science) according to the manufacturer's instruction. Myosin II ATPase inhibitor (-)-blebbistatin (Wako) was used at 10 μM concentration, and actin polymerization inhibitor Latrunculin A (Wako) was used at 10 nM concentration. An equivalent amount of dimethyl sulfoxide (DMSO) was administered to cells as vehicle control.
Dulbecco's Modified Eagle Medium (Wako) containing 10% (v/v) heat-inactivated fetal bovine serum (SAFC Biosciences) and 1% penicillin-streptomycin (Wako) in a humidified 5% CO2 incubator at 37℃. Expression plasmids encoding mClover2-tagged β-actin were constructed by inserting a human β-actin gene, which was digested with XhoI and BamHI restriction enzymes from the EYFP-actin vector (#6902-1, Clontech) into the mClover2-C1 vector (Addgene plasmid #54577, a gift from Michael Davidson). The plasmids were transfected to cells using Lipofectamin LTX with Plus Reagent (Thermo Fischer Science) according to the manufacturer's instruction. Myosin II ATPase inhibitor (-)-blebbistatin (Wako) was used at 10 μM concentration, and actin polymerization inhibitor Latrunculin A (Wako) was used at 10 nM concentration. An equivalent amount of dimethyl sulfoxide (DMSO) was administered to cells as vehicle control.
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