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. 2016 Jun 6;37(5):428–443. doi: 10.1016/j.devcel.2016.05.003

Figure 2.

Figure 2

The EPS15 μHD Interaction Motif In Vivo

(A and B) Representative single confocal optical sections of basal surface of HeLa SS6 cells transfected with either Tac-FCHO1 μHD (A) or Tac (B) as indicated. Fixed cells were stained with anti-Tac or anti-EPS15 antibodies. Enlarged color-separated regions corresponding to the boxed areas are shown and Golgi-localized Tac and EPS15 (arrows) are indicated. Scale bar, 10 μm.

(C–E′) Typical basal confocal sections of fixed HeLa cells double transfected with the indicated Tac and tdRFP-EPS15 plasmids and stained with antibodies against Tac and GPP130 as indicated (C′–E′). Corresponding medial optical sections of the boxed regions are shown (C–E) indicating the Golgi pool (arrowheads) of Tac protein. Scale bar, 10 μm.

(F and G) Representative medial confocal optical sections of HeLa cells transiently transfected with Tac-FCHO1 μHD and either tdRFP-EPS15 (1–636) or (1–614) as indicated after fixation and staining with anti-Tac and anti-Eps15 antibodies. Enlargements of boxes as in (A). Note that the anti-Eps15 is a peptide-specific antibody raised against the extreme C terminus of the protein, which is truncated in the two tagged exogenous proteins. Scale bar, 10 μm.