Figure 4
Evaluation of binding and internalization of (Rap) 2 -E1-(Rap) 2 in MDA-MB-468. A, flow cytometric analyses showing equivalence in binding of (Rap)2-E1-(Rap)2 and hRS7 IgG to MDA-MB-468 at three concentrations (1, 5 and 25 nM). The CD20-targeting (Rap)2-20-(Rap)2 and hA20 served as negative controls; B, following incubation, pHĀ 2.5 treatment, and fixation as described in the Methods, cells were stained with GAH- Alexa Fluor 488 (green) to detect intracellular (Rap)2-E1-(Rap)2, and also with Hoechst 33258 (blue) and human transferrin-conjugated Alexa Fluor 568 (red) to reveal the nucleus and endosomes, respectively.