NPC-039 and NPC-BM cells were seeded into a 6-well plate at a density of 3 × 105 cells per well and exposed to the indicated dose of picrasidine I (0, 10, 20, 40 µM). Subsequently, cells were collected and stained with Muse™ Caspase 7-AAD reagent and Muse™ Caspase-3/7 reagent (Luminex). The protocol was conducted according to the Muse™ Caspase-3/7 Kit user’s guide. The percentage of active caspase-3/7 cells was measured using the Muse cell analyzer flow cytometer (MilliporeSigma) and analyzed using Muse Cell Soft V1.4.0.0 Analyzer Assays (MilliporeSigma).
Muse cell soft v1.4.0.0 analyzer assays
Manufactured by Merck Group
Sourced in United States
The Muse Cell Soft V1.4.0.0 Analyzer Assays is a laboratory equipment product from Merck Group. It is designed to analyze cell samples using soft flow cytometry technology.
Automatically generated - may contain errors
9 protocols using muse cell soft v1.4.0.0 analyzer assays
1
Apoptosis Induction Assay for NPC Cells
2
Mitochondrial Membrane Potential Analysis
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Mitochondrial membrane potential analysis has previously been described [57 (link)]. Briefly, cells that were treated with the indicated dose of picrasidine I were collected and stained with Muse MitoPotential dye (Luminex, Austin, TX, USA). The stained cells were measured using the Muse cell analyzer flow cytometer (MilliporeSigma, Burlington, MA, USA) and analyzed using Muse Cell Soft V1.4.0.0 Analyzer Assays (MilliporeSigma).
3
Mitochondrial Potential and Viability Analysis
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As previously described,65 (link) cells were harvested and collected (300 × g, 5 min). Treatment cell precipitates were stained with Muse MitoPotential dye (20 min, 37°C), and then 7-AAD was added for 5 min. The experiment signals were analyzed by the Muse Cell Soft V1.4.0.0 Analyzer Assays (Millipore).
4
Annexin V Apoptosis Assay
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As previously described,66 cells (1 × 105) were harvested and suspended in 100 μL PBS with 2% BSA after treatment. Cells were then stained with Muse Annexin V & Dead Cell Reagent (100 μL) for 20 min at room temperature in the dark. The signals were analyzed by the Muse Cell Soft V1.4.0.0 Analyzer Assays (Millipore).
5
Cell Cycle Analysis of Chrysosplenol D
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Cells treated with the indicated doses of chrysosplenol D (0, 25, 50, and 100 µM) were collected and fixed with 70% ethanol for 24 h at −20 °C. After discarding ethanol, we incubated cells with the Muse cell cycle reagent (Merck Millipore, Burlington, MA, USA) for 30 min in the dark. Subsequently, cell cycle distribution was measured using a Muse cell analyzer flow cytometer (Merck Millipore), and data were analyzed using Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
6
Quantifying Cellular Oxidative Stress
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The user guide of the Muse Oxidative Stress Kit (Cat. No. MCH100111, Merck Millipore) describes the oxidative stress detection method. First, the cells were planted in a 6-well plate (1×104 cells/well) and incubated with various DC concentrations for 24 h as previously described (23 (link)). The collected cells were processed under conditions previously studied (23 (link)). The obtained cells were added to the Muse Oxidative Stress working solution reagent reaction at 37°C for 30 min, and the results were analyzed using Muse Cell Analyzer flow cytometry and the data were analyzed using the Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
7
Evaluating Mitochondrial Potential in Cells
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First, the cells were planted in a 6-well plate (1×104 cells/well) and incubated with various DC concentrations for 24 h as previously described (23 (link)). The collected cells were processed under conditions previously studied (23 (link)). The obtained cells were added to Muse Mitopotential Assay Kit (cat. no. MCH100110, Merck Millipore) reaction, and the results were analyzed using Muse Cell Analyzer flow cytometry and the data were analyzed using Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
8
Cell Viability Assay for Drug Screening
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Cell viability was determined following methods described previously (22 (link)). The cells (1×104 cells/well) were cultured in each well for 12 h and further treated with various DC concentrations for 24 h. These cells were collected and suspended in phosphate-buffered saline (PBS) followed by incubation with reagents contained in the Muse Annexin V and Dead Cell Kit (cat. no. MCH100105; Merck Millipore) in the dark at room temperature. Results were analyzed using Muse Cell Analyzer flow cytometry (Merck Millipore) and the data were analyzed using Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
9
Caspase-3/7 Detection in Cell Lines
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The analysis was performed as previously described (24 (link)). The user guide of the Muse Caspase-3/7 Kit (cat. no. MCH100108, Merck Millipore) describes the caspase-3/7 detection method. After processing the DC, the cells were obtained and stained with the reagent of Muse Caspase-3/7. The experimental results were detected using a flow cytometer and analyzed using Muse Cell Analyzer flow cytometry and the data were analyzed using the Muse Cell Soft V1.4.0.0 Analyzer Assays (Merck Millipore).
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