Figure 3.

The Chi-Tn mAb is internalized into cancer cells. (A) Jurkat, Shin-3 or TA3Ha cells were incubated for 15 min on ice with the Chi-Tn mAb or with a control antibody (IvIg for human cells or trastuzumab for murine cells) at 20 µg/mL, washed, then transferred to 37°C for the indicated times. Cells were then labeled with GaH-Fc-PE, and the Chi-Tn mAb M.F.I. was determined by flow cytometry in the DAPI-negative living cells gate. For each sample, the Chi-Tn M.F.I. is expressed as a percentage of the Chi-Tn M.F.I. obtained for cells not transferred to 37°C (control cells). (B) Jurkat, OvCar-3, Shin-3 or TA3Ha cells were incubated on ice with the Chi-Tn mAb for 15 min, washed and transferred to 37°C for the indicated period of time. Cells were then fixed to glass coverslips and labeled. Yellow: actin network; pink: membrane-bound or internalized Chi-Tn mAb; blue: DAPI. Arrows indicate examples of internalized Chi-Tn mAb.