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. Author manuscript; available in PMC: 2012 Jun 25.
Published in final edited form as: Biochem J. 2009 Jul 29;422(1):43–52. doi: 10.1042/BJ20081189

Figure 3. The short ZnT2 isoform is localized to the plasma membrane in mammary epithelial cells.

Figure 3

(A) Representative immunoblot (IB) of crude membrane proteins (100 μg protein/lane) from HC11 cells transfected with pcDNA3.1 (lane 1, mock-transfected control) or cells expressing the 35 kDa ZnT2 short–HA (lanes 2–3) detected with HA antibody. (B) Confocal micrographs of cells transfected with pcDNA3.1-shortHA, to express ZnT2 short–HA, detected with HA antibody and visualized with Alexa Fluor® 488-conjugated anti-mouse IgG. Confocal micrographs illustrate detection of the ZnT2 short-HA at the cell periphery (arrows); COX IV (cytochrome c oxidase subunit IV; red) was utilized for spatial orientation. Asterisks represent individual transfected cells (n = 5 cells expressing ZnT2 short-HA). Bar, 10 μm. (C) Plasma membrane association was verified by immunoblot of cell surface proteins isolated from cells expressing the 35 kDa ZnT2 short-HA, captured with Ultralink Neutravidin, detected with HA antibody. Non-specific binding to avidinated beads was assessed in proteins captured from cells not exposed to biotin (—) and compared with biotinylated cell surface proteins.