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. 2016 Mar 23;7(18):25443–25460. doi: 10.18632/oncotarget.8327

Figure 3. Localization studies of ERM proteins, Ebp50 and ErbB2 in SKBR3 cells and protein proximity analysis.

Figure 3

(A) Confocal microscopy of ERM proteins and Ebp50. Untreated cells were fixed, permeabilized and stained by specific antibodies for endogenous levels of ezrin, radixin and Ebp50 (max. projection). In SKBR3 cells ezrin and radixin strongly colocalizes with Ebp50 at the plasma membrane. (B) Confocal microscopy (single plane section) and (C) 3D-SIM (max. projections) of ErbB2 and Ebp50 localization. In untreated control conditions, ErbB2 shows a strong colocalization with Ebp50 in lamellipodia-like structures at the plasma membrane. (D-F) Analysis of protein proximity by PLA experiments. SKBR3 cells were transfected either with non-targeting control siRNA or ezrin/radixin specific siRNA. Cells were fixed 72 h after transfection and PLA experiments were performed. The depletion of ezrin or radixin strongly reduces the interaction of ErbB2 with Ebp50 (D), Hsp90 (E) and ErbB3 (F). All results are shown as mean +/− SEM (*P < 0.05; ***P < 0.001). Scale bars: 10 μm (A, B), 8 μm (C).