Obtained human cardiomyocytes were seeded on a flexible surface (BF‐3001C, FlexCell) subjected to 15% equiaxial mechanical stretch at 1 Hz for 24 h or 48 h using a FX‐4000 system (FlexCell) as mentioned previously.23 Cardiomyocytes were cultured in anaerobic pouches (260 683, BD) for 72 h to create hypoxic conditions. Additionally, cardiomyocytes were incubated with 100 ng/mL tumour necrosis factor alpha (TNFα; SRP3177, Sigma‐Aldrich) for 72 h in order to mimic an inflammatory environment. To inhibit lysosomal exocytosis, hPSC‐CM were incubated with the small molecule vacuolin‐1 (20 425, Cayman Chemical) at a concentration of 1 μM in the cell medium.24
Vacuolin 1
Manufactured by Cayman Chemical
Vacuolin-1 is a chemical compound that functions as a selective inhibitor of the vacuolar ATPase (V-ATPase) enzyme. V-ATPase is responsible for the acidification of intracellular compartments in eukaryotic cells. Vacuolin-1 is commonly used as a research tool to study the role of V-ATPase in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anaerobic pouches, by BD (1 mentions)
Fx 4000 system, by Flexcell (1 mentions)
Tnf α, by Merck Group (1 mentions)
Bf 3001c, by Flexcell (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Retinoic acid, by Merck Group (1 mentions)
Sb202190, by Cayman Chemical (1 mentions)
Apilimod, by Cayman Chemical (1 mentions)
Ly2228820, by Cayman Chemical (1 mentions)
Ym201636, by Cayman Chemical (1 mentions)
Skepinone l, by Cayman Chemical (1 mentions)
Birb796, by Cayman Chemical (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
3 protocols using vacuolin 1
1
Cardiac Stress and Inflammation Protocol
2
HL-60 Cell Culture and Treatment
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HL-60 human myeloblastic leukemia cells were a generous gift of Dr. Robert Gallagher. They are cultured in RPMI 1640 supplemented with 5% heat-inactivated fetal bovine serum (GE Healthcare, Chicago, IL) and 1% antibiotic/antimycotic (Thermo Fisher Scientific, Waltham, MA) in a 5% CO2 humidified atmosphere at 37 °C as previously described [34 (link)]. Retinoic acid (Sigma, St. Louis, MO) was added from a stock of 5 mM in ethanol to make a final concentration of 1 µM. Vacuolin-1 (Cayman Chemical, Ann Arbor, MI) was added from a stock of 0.25 mM in DMSO to make a final concentration of 0.25 µM. Viability assessed by 0.1% trypan blue exclusion was routinely in excess of 95% in all cultures except toxicity experiments in supplemental data that used vacuolin in excess of 2.5 µM where a sub-G1 population was also apparent.
3
Compound Preparation for Cell Culture
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