Cells were scraped from the culture dish and then washed three times with cold PBS. Staining was performed according to the manufacturer’s recommended protocol using PE anti-mouse CD86 (12-0862-82, Thermo Fisher, USA) and APC anti-mouse CD206 (17–2061-82, Thermo Fisher, USA) antibodies. Data acquisition and further analysis was conducted using the CytExpert software (version 10.0.7; Tree Star, Ashland, OR, USA).
Cytexpert software
Manufactured by Tree Star
Sourced in United States
CytExpert is a software application designed for the analysis and management of flow cytometry data. It provides tools for data visualization, gating, and statistical analysis of cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Apc anti mouse cd206, by Thermo Fisher Scientific (1 mentions)
Cd11b fitc, by Thermo Fisher Scientific (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
Aldehyde sulphate latex beads, by Thermo Fisher Scientific (1 mentions)
Beckman cytoflex, by Beckman Coulter (1 mentions)
Cl488 66699, by Proteintech (1 mentions)
Facsaria 2, by BD (1 mentions)
3 protocols using cytexpert software
1
Macrophage Surface Marker Analysis
2
Liver Leukocyte Isolation and CD11b Analysis
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Liver leukocytes of M. fortis were prepared by Percoll gradient centrifugation after being dispersed through a 70-μm nylon strainer. The cell precipitates were washed twice and suspended in PBS containing 0.1% BSA and 0.05% sodium azide. A total of 2 × 106 cells per 100 μL were incubated with CD11b-FITC (eBioscience, CA) for 30 min at 4 °C in the dark. The expression of CD11b was analyzed on a CytoFLEX flow cytometer (Beckman Coulter, USA) using CytExpert software (Treestar, CA).
3
Quantifying ACE2 Expression on Cells and EVs
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To detect the level of ACE2 on cell membrane surface, cells were prepared according to standard protocols and suspended in 2% FBS with PBS. Anti‐human ACE2 antibody (CL488‐66699, Proteintech) was used for ACE2 detection. To detect the level of ACE2+ EV, EVs were incubated with aldehyde/sulphate latex beads (4 µm, Invitrogen, no.1743119) in PBS for 1h at 4 °C. EV‐bound beads were incubated with anti‐ACE2 FITC‐conjugated antibody (CL488‐66699, Proteintech) for 30 min. The percentage of positive beads referred to as the percentage of ACE2+ EVs was calculated relative to the total number of beads analyzed per sample. All samples were analyzed with Beckman CytoFlex (Beckman) or FACSAria II (Becton Dickinson) machines. FACS data were analyzed with CytExpert software and FlowJo (TreeStar).
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