Figure 3. Construction of tagged T-cadherin. (A) Prediction of T-cadherin structure, showing insertion of schematic 3xGly-mRFP-3xGly peptide folding separately in between cadherin motifs four and five. (B) Western blot of CHO cells with or without transfection with wild type T-cadherin or the Tcad-mRFP construct. (C–E) T-cadherin (green) immunostaining performed on living non-permeabilized cells, illustrating Tcad surface localization. Tcad-mRFP fluorescence-red. Scale bar: 10 μm. (C) Wild type CHO cells. (D) CHO cells transfected with Tcad-mRFP. (E) CHO cells transfected with wild type T-cadherin. Note that some Tcad-mRFP is detected in the cytoplasm (D), which probably reflects protein during processing and transport.