Anti cd71 apc m a712

Flow cytometry analysis was done as described earlier^{6 (link)}. Cells (1 × 10⁵) obtained after enzymatic dissociation of 1° and 2° cervicospheres were stained with anti-CD49f-FITC (GoH3; BD-Pharmingen), anti-CD71-APC (M-A712; BD-Pharmingen), and/or anti-CD133-PE for 60 minutes at 4 °C in staining buffer (2% BSA in 185 PBS). Corresponding isotypes were used as control. Cells were washed in PBS, centrifuged, and finally resuspended in 300 µL analysis buffer (1% BSA/2 mmol/L EDTA in PBS). FACS sorting was done as described earlier^{6 (link)}. In brief, 1 × 10⁶ cells were incubated with anti-CD49f-FITC, anti-CD71-APC, and/or anti-CD133-PE, and were finally resuspended in 500 µL analysis buffer. The dead cells and debris were excluded after 7-AAD staining (Invitrogen). Data were collected on FACSAriaIII cell sorter (BD Biosciences) and analyzed using FlowJo software (TreeStar).