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. 2016 Jul 28;20(10):1999–2011. doi: 10.1111/jcmm.12899

Figure 6.

Figure 6

Antibody‐induced internalization of HER2 depends on dynamin 2 but only partially on clathrin. (A) OE19 cells were transiently transfected to express GFP‐tagged dynamin 2 K44A (Dyn2‐K44A). Cells were treated with the mAb mixture in MEM for 4 hrs, before fixation, permeabilization and immunostaining for human IgG. Micrographs representative for three experiments are shown. Left panel; overlay of Dyn2‐K44A (green) and human IgG. Right panel; human IgG only. Insets showing high magnification of the framed areas demonstrate lack of internalization in cells expressing dynamin 2 K44A (green frame), scale bar: 10 μm. (B) Clathrin heavy chain (CHC) was knocked down in OE19 cells by siRNA as described in Materials and Methods. Non‐targeting (NT) siRNA was used as control. Cells were incubated in MEM with the mAb mixture for 4 hrs before fixation. Upon permeabilization, the cells were immunostained using antibodies to human IgG. Insets show high magnification of the corresponding framed area in each panel. Micrographs representative for three experiments are shown, scale bars: 15 μm.