Fitc conjugated anti cd24 antibody

Cells were collected and washed twice with permeabilization wash buffer and were then resuspended in flow cytometry staining buffer (Bio-Rad), and cell numbers were counted. The dissociated cells were then stained with APC-conjugated anti-CD44 antibody (1:50, Cat# 559942, BD Pharmingen) and FITC-conjugated anti-CD24 antibody (1:50, Cat# 555427, BD Pharmingen) for 30 min at 4 °C. After washing twice in flow cytometry buffer, the cells were analyzed by a FACSCalibur flow cytometer (BD Biosciences). For the ALDEFLUOR Assay, cells were harvested by trypsinization, washed in PBS, labeled with Aldefluor Reagent (StemCell Technologies, Grenoble, France) and incubated at 37 °C for 45 min. Finally, all samples were analyzed by a FACS machine (BD Calibur).

PTX-EXO and/or si-circBACH1 treated cells in the presence of PTX were dual-labeled with a APC-conjugated anti-CD44 antibody and an FITC-conjugated anti-CD24 antibody (BD Pharmingen, San Diego). 1 × 10⁵ cells were labeled and incubated with FITC-conjugated anti-CD24 and APC-conjugated anti-CD44 antibody at 4 °C in the dark for 1 h. The cells were washed and then analyzed by flow cytometer (BD FACS Aria II).