Anti human cd24 pb

Cells were lifted with Accutase® (Corning, Catalog # 25-058-CI) and stained with anti-human CD44-FITC (BD Pharmingen) and anti-human CD24-PB (Exbio) antibodies at manufacturer’s recommended dilutions in 0.1% BSA (Bovine serum albumin, Sigma) diluted in DPBS for 1 h in a rotary shaker at room temperature. Cells were analysed in the presence of propidium iodide (1 µg/mL) using a BD LSRFortessa (BD Biosciences). After doublet discrimination and compensation for spectral overlap, data were analysed by using FlowJo Software (BD Biosciences). For TGFβR2 surface expression, cells were stained with primary antibody (RandD Systems, Cat# AF-241-NA) as per manufacture recommended dilutions for 1 h and then with secondary goat antibody for 1 h.

Cells were harvested with Accutase® (Corning, Catalog # 25-058-CI) and stained with anti-human CD44-FITC (BD Pharmingen), anti-human CD24-PB (Exbio) and anti-human EpCAM-APC (Biolegend) antibodies, as per manufacturer-recommended dilutions for 1 h at room temperature on a rotary shaker. Cells were analyzed in the presence of propidium iodide (1 µg/mL) using a BD LSR Fortessa (BD Biosciences). After doublet discrimination and compensation for spectral overlap, samples were analyzed using FlowJo Software v10.0.7 (BD Biosciences). For sorting, anti-human EpCAM-PerCP/Cy5.5 (Biolegend) antibody was used and cells were sorted using a BD FACS Aria IIu sorter (BD Biosciences).