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. 2013 Jun;141(6):691–703. doi: 10.1085/jgp.201210937

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© 2013 Ponissery Saidu et al.

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Figure 5. — Localization of ANO2 Isoform B and Isoform B_Δ4 in HEK cells. (A) Immunofluorescence images of HEK293T cells transfected with either ANO2 Isoform B::GFP (top row) or Isoform B_Δ4::GFP (bottom row). The left panels show Isoform B::GFP or B_Δ4::GFP staining. Biotin labeling, shown in the middle panels, is used as a membrane marker. The right panels show merged images with DAPI as a nuclear marker. White arrowheads point to areas of Isoform B::GFP or B_Δ4::GFP colocalization with biotin. (B) Immunogold labeling against GFP in HEK293T cells. TrkA::GFP is used as a membrane localization control (bottom left panel), and GFP is used as a cytoplasmic control (bottom right panel). Black arrows point to the cell membrane. White arrowheads point to several immunogold particles. PM, cell membrane; cyto, cytoplasm. The GFP panel shows two adjacent cells: one expressing a high level of GFP (GFP+) and the other expressing little or no GFP (GFP−). The black dashed lines demarcate the GFP+ and the GFP− cell. For quantification of the spatial distribution of gold particles, see Fig. S3.