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Fig. 3 | BMC Biology

Fig. 3

From: A straightforward approach for bioorthogonal labeling of proteins and organelles in live mammalian cells, using a short peptide tag

Fig. 3

Plasma membrane (PM) labeling obtained via the minimal GCE-tag is comparable to GFP and site-specific labeling. HEK293T (a) or COS7 (b–e) cells were transfected with pBUD-Pyl-RS plasmids carrying GCE-tag-GFP-CAAX, GFP-CAAX mutated at position 150 to TAG or a WT version of GFP-CAAX, and incubated for 48 h in the presence of BCN-Lys. Cells were then subjected to western blot analysis (a) or labeled with SiR-Tet for 1 h and imaged live (b–e). Images represent single confocal slices of representative cells. d Line intensity profiles obtained in cells expressing the indicated constructs and labeled with SiR-Tet. Upper panel: zoomed-in images of confocal slices. Bottom panel: intensity values along the dashed line drawn in the images. e SNR values measured for GFP and SiR in cells expressing the different versions of GFP-CAAX. n = 30. Results were obtained in at least 3 independent experiments. Scale bars: b, c 10 μm, d 2 μm

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