Figure 5

Fluorescence imaging of mTFP1 fusion constructs. (A)-(K) N-terminal fusion constructs; for each fusion protein the linker amino acid length is indicated after the name of the targeted organelle or fusion protein: (A) mTFP1-α-actinin-19 (human non-muscle); (B) mTFP1-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mTFP1-Cx43-7 (rat α-1 connexin-43); (D) mTFP1-keratin-17 (human cytokeratin 18); (E) mTFP1-endoplasmic reticulum-3 (calreticulin signal sequence (51 nucleotides) and KDEL retention sequence); (F) mTFP1-paxillin-22 (chicken); (G) mTFP1-EB3-7 (human microtubule-associated protein; RP/EB family); (H) mTFP1-lysosomes-20 (rat lysosomal membrane glycoprotein 1); (I) mTFP1-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (J) mTFP1-vimentin-7 (human); (K) mTFP1-zyxin-7 (human). (L)-(T) C-terminal fusion constructs: (L) mTFP1-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (M) mTFP1-lamin B1–10 (human); (N) mTFP1-β-actin-7; (O) mTFP1-clathrin light chain-15 (human); (P) mTFP1-fibrillarin-7 (human); (Q) mTFP1-vinculin-23 (human); (R) mTFP1-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S)mTFP1-β-tubulin-6 (human); (T) mTFP1-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras). The cell line used for expressing mTFP1 fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (A), (G), (K), (N) and Q) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.