Quantification of RAW264.7 Cell Phenotypes

RAW264.7 (1 × 10⁴) cells cultured with HA and β‐TCP samples for 2 and 5 days were collected and then respectively incubated with peridinin chlorophyll protein (Percp, BioLegend, USA)‐conjugated anti‐mouse CD11c and activated protein C (APC, eBioscience, USA)‐conjugated anti‐mouse CD206 at room temperature for 30 min without light after fixation, membrane breaking and blocked with CD16/32 (BD pharmingen) steps. Finally, the cells were resuspended in 200 µL PBS and subjected to FACSCalibur flow cytometry (Beckman, CytoFLEXS, USA).

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Sun X., Li Z., Wang X., He J, & Wu Y. (2024). Inorganic Phosphate as “Bioenergetic Messenger” Triggers M2‐Type Macrophage Polarization. Advanced Science, 11(13), 2306062.

Publication 2024

Percp CD16/32 FACSCalibur flow cytometry

Corresponding Organization : Sichuan University

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Variable analysis

independent variables

HA (hyaluronic acid) samples
β‐TCP (beta‐tricalcium phosphate) samples

dependent variables

Expression of CD11c (a marker for dendritic cells)
Expression of CD206 (a marker for M2 macrophages)

control variables

RAW264.7 cells (a murine macrophage cell line)
Cell culture duration (2 days and 5 days)

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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