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TaKaRa human h2b
Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); <t>(I)mWasabi-H2B-6</t> (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
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Images

1) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

Live cell imaging of mTFP1 fusion vectors . (A)-(D) Laser scanning confocal images of a single HeLa cell expressing mTFP1-H2B-6 (N-terminus; human) progressing through prophase, metaphase, anaphase and interphase, respectively. (E)-(H) Spinning disk confocal images selected from a time-lapse series of HeLa cells expressing mTFP1-annexin (A4)-12 (C-terminus; human) during ionomycin-induced translocation to the plasma and nuclear membranes [35]: (E) 0 min, ionomycin added; (F) 5 min; (G) 7 min; (H) 9 min. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mTFP1 fusion vectors . (A)-(D) Laser scanning confocal images of a single HeLa cell expressing mTFP1-H2B-6 (N-terminus; human) progressing through prophase, metaphase, anaphase and interphase, respectively. (E)-(H) Spinning disk confocal images selected from a time-lapse series of HeLa cells expressing mTFP1-annexin (A4)-12 (C-terminus; human) during ionomycin-induced translocation to the plasma and nuclear membranes [35]: (E) 0 min, ionomycin added; (F) 5 min; (G) 7 min; (H) 9 min. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Expressing, Translocation Assay

2) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

3) Product Images from "TEMPERATURE DEPENDENT PHASE BEHAVIOR AND PROTEIN PARTITIONING IN GIANT PLASMA MEMBRANE VESICLES"

Article Title: TEMPERATURE DEPENDENT PHASE BEHAVIOR AND PROTEIN PARTITIONING IN GIANT PLASMA MEMBRANE VESICLES

Journal: Biochimica et biophysica acta

doi: 10.1016/j.bbamem.2010.03.009

Partitioning of wt Fyn anchor eGFP construct. (a) Representative fluorescence images comparing protein (green, left) and lipid dye (red, right) fluorescence in a phase separated HeLa cell GPMV. Scale bars, 2 µm. (b) Fluorescence intensity ratio distribution for protein and lipid dye demonstrates primarily disordered phase partitioning of the wt Fyn protein anchor. Scale bars, 2 µm.
Figure Legend Snippet: Partitioning of wt Fyn anchor eGFP construct. (a) Representative fluorescence images comparing protein (green, left) and lipid dye (red, right) fluorescence in a phase separated HeLa cell GPMV. Scale bars, 2 µm. (b) Fluorescence intensity ratio distribution for protein and lipid dye demonstrates primarily disordered phase partitioning of the wt Fyn protein anchor. Scale bars, 2 µm.

Techniques Used: Construct, Fluorescence

4) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

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.
Figure Legend Snippet: 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.

Techniques Used: Fluorescence, Imaging, Construct, Sequencing, Expressing

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

5) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

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.
Figure Legend Snippet: 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.

Techniques Used: Fluorescence, Imaging, Construct, Sequencing, Expressing

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

6) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

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.
Figure Legend Snippet: 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.

Techniques Used: Fluorescence, Imaging, Construct, Sequencing, Expressing

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

7) Product Images from "Voltage-dependent Dynamic FRET Signals from the Transverse Tubules in Mammalian Skeletal Muscle Fibers"

Article Title: Voltage-dependent Dynamic FRET Signals from the Transverse Tubules in Mammalian Skeletal Muscle Fibers

Journal: The Journal of General Physiology

doi: 10.1085/jgp.200709831

Membrane expression of EGFP-F and ECFP-F in FDB muscle fibers. (A) TPLSM image section obtained at the medial plane of a muscle fiber expressing EGFP-F. n denotes nucleus; arrowheads point to the cytoplasmic side of a nucleus; arrows point toward areas of increased fluorescence close to the poles of nuclei. (B) Fluorescence intensity profile obtained from the area delimited by the white rectangle in A. (C and D) TPLSM image sections of an FDB muscle transfected with pECFP-F and stained extracellularly with di-8-ANEPPS. ECFP-F and di-8-ANEPPS fluorescence images are shown in C and D, respectively. (E) Superimposition of image sections in C and D. (F) Normalized fluorescence intensity profiles measured from the area delimited by the rectangles in Fig. 2, C (cyan trace) and D (red trace). The vertical calibration bar is 20 μm and applies to all the images.
Figure Legend Snippet: Membrane expression of EGFP-F and ECFP-F in FDB muscle fibers. (A) TPLSM image section obtained at the medial plane of a muscle fiber expressing EGFP-F. n denotes nucleus; arrowheads point to the cytoplasmic side of a nucleus; arrows point toward areas of increased fluorescence close to the poles of nuclei. (B) Fluorescence intensity profile obtained from the area delimited by the white rectangle in A. (C and D) TPLSM image sections of an FDB muscle transfected with pECFP-F and stained extracellularly with di-8-ANEPPS. ECFP-F and di-8-ANEPPS fluorescence images are shown in C and D, respectively. (E) Superimposition of image sections in C and D. (F) Normalized fluorescence intensity profiles measured from the area delimited by the rectangles in Fig. 2, C (cyan trace) and D (red trace). The vertical calibration bar is 20 μm and applies to all the images.

Techniques Used: Expressing, Fluorescence, Transfection, Staining

Effects of DPA on the electrical and optical properties of a voltage-clamped skeletal muscle fiber expressing EGFP-F. The fiber was clamped at an HP = −90 mV and stimulated with depolarizing and hyperpolarizing voltage pulses of 40, 60, 80, and 100 mV. Data in A and B were obtained before and after DPA staining, respectively. (A) Top panels show capacitive currents in response to 10-ms depolarizing (left) or hyperpolarizing (right) pulses. Bottom panel shows, in an expanded ordinate scale, the corresponding ECFP-F fluorescence records. (B) Capacitive (top) currents and QRET transients (bottom) in response to 20-ms depolarizing (left) and hyperpolarizing (right) voltage-clamp pulses from the same fiber in A, after DPA staining. The rising phase of QRET transients elicited by depolarizing pulses could be adjusted to single exponential functions with τ = 0.4, 1.1, 1.1, and 1.2 ms for the black, red, green, and blue traces, respectively.
Figure Legend Snippet: Effects of DPA on the electrical and optical properties of a voltage-clamped skeletal muscle fiber expressing EGFP-F. The fiber was clamped at an HP = −90 mV and stimulated with depolarizing and hyperpolarizing voltage pulses of 40, 60, 80, and 100 mV. Data in A and B were obtained before and after DPA staining, respectively. (A) Top panels show capacitive currents in response to 10-ms depolarizing (left) or hyperpolarizing (right) pulses. Bottom panel shows, in an expanded ordinate scale, the corresponding ECFP-F fluorescence records. (B) Capacitive (top) currents and QRET transients (bottom) in response to 20-ms depolarizing (left) and hyperpolarizing (right) voltage-clamp pulses from the same fiber in A, after DPA staining. The rising phase of QRET transients elicited by depolarizing pulses could be adjusted to single exponential functions with τ = 0.4, 1.1, 1.1, and 1.2 ms for the black, red, green, and blue traces, respectively.

Techniques Used: Expressing, Staining, Mass Spectrometry, Fluorescence

QRET transient amplitude and DPA//EGFP-F and DPA//ECFP-F spectral overlap. (A) Spectral overlap between DPA absorption spectrum (red trace) and EGFP-F normalized emission spectrum (green trace). (B) Spectral overlap between DPA absorption spectrum (red trace) and ECFP-F normalized emission spectra (cyan trace). EGFP-F and ECFP-F emission spectra were determined in vivo and DPA absorption spectrum was measure in vitro. (C) Peak amplitude of EGFP-F (green hatched bar) and ECFP-F (cyan hatched bar) QRET transients evoked by AP stimulation. The averages values (in −ΔF/F) were calculated from 6 and 12 fibers for EGFP-F and ECFP-F, respectively. The error bar is the SEM. The asterisk indicates statistical significance (P
Figure Legend Snippet: QRET transient amplitude and DPA//EGFP-F and DPA//ECFP-F spectral overlap. (A) Spectral overlap between DPA absorption spectrum (red trace) and EGFP-F normalized emission spectrum (green trace). (B) Spectral overlap between DPA absorption spectrum (red trace) and ECFP-F normalized emission spectra (cyan trace). EGFP-F and ECFP-F emission spectra were determined in vivo and DPA absorption spectrum was measure in vitro. (C) Peak amplitude of EGFP-F (green hatched bar) and ECFP-F (cyan hatched bar) QRET transients evoked by AP stimulation. The averages values (in −ΔF/F) were calculated from 6 and 12 fibers for EGFP-F and ECFP-F, respectively. The error bar is the SEM. The asterisk indicates statistical significance (P

Techniques Used: In Vivo, In Vitro

8) Product Images from "Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein"

Article Title: Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein

Journal: Nature methods

doi: 10.1038/nmeth.2888

Structural basis of red-shifting in mCardinal. (a) Structure of Neptune showing the water molecule hydrogen-bonded to the acylimine oxygen of the chromophore. The fluorescent protein is viewed down the axis of the beta-barrel from the direction of the N- and C-termini. Beta-strands and alpha-helices are depicted in cartoon form. The chromophore, the Met63 side chain attached to the chromophore, and the water are depicted in stick representation with carbon colored lavender, nitrogen blue, and oxygen red. Numbers indicate distances between the water oxygen atom and hydrogen-bonding partners. Cyan labels indicate positions mutated to amino acids with side chains capable of donating hydrogen bonds to the acylimine oxygen: Met11, Leu13, Ser28 and Gly41. (b,c) Hydrogen bond interactions between Thr28, Gln41, and the chromophore acylimine in mCardinal (b) or mCardinal-V218E (c) . Rendering is of chain A. Distances shown are averages of measurements from chains with temperature factors for Gln41 amide atoms lower than those for the hydrogen-bonding water molecule in the Neptune structure, specifically chains A and B for mCardinal and chains A and C for mCardinal-V218E. Mesh depicts electron density in the Fo-Fc OMIT map contoured to 3.0 sigma within 2.0 Å of the atoms displayed in stick form.
Figure Legend Snippet: Structural basis of red-shifting in mCardinal. (a) Structure of Neptune showing the water molecule hydrogen-bonded to the acylimine oxygen of the chromophore. The fluorescent protein is viewed down the axis of the beta-barrel from the direction of the N- and C-termini. Beta-strands and alpha-helices are depicted in cartoon form. The chromophore, the Met63 side chain attached to the chromophore, and the water are depicted in stick representation with carbon colored lavender, nitrogen blue, and oxygen red. Numbers indicate distances between the water oxygen atom and hydrogen-bonding partners. Cyan labels indicate positions mutated to amino acids with side chains capable of donating hydrogen bonds to the acylimine oxygen: Met11, Leu13, Ser28 and Gly41. (b,c) Hydrogen bond interactions between Thr28, Gln41, and the chromophore acylimine in mCardinal (b) or mCardinal-V218E (c) . Rendering is of chain A. Distances shown are averages of measurements from chains with temperature factors for Gln41 amide atoms lower than those for the hydrogen-bonding water molecule in the Neptune structure, specifically chains A and B for mCardinal and chains A and C for mCardinal-V218E. Mesh depicts electron density in the Fo-Fc OMIT map contoured to 3.0 sigma within 2.0 Å of the atoms displayed in stick form.

Techniques Used:

Comparison of mCardinal with mNeptune1, iRFP and Clover GFP for non-invasive visualization of muscle regeneration in living mice. (a) Representative fluorescence images of tibialis anterior (TA) muscles injected with 1 million myoblasts expressing iRFP and mCardinal or iRFP and mNeptune1. Images were taken in the absence of exogenous biliverdin. The bar graph shows contrast over background (mean ± SEM, n = 6 and n = 8 for mNeptune1 and mCardinal, respectively). Asterisks indicate significant differences by unpaired two-tailed Student t test (* P
Figure Legend Snippet: Comparison of mCardinal with mNeptune1, iRFP and Clover GFP for non-invasive visualization of muscle regeneration in living mice. (a) Representative fluorescence images of tibialis anterior (TA) muscles injected with 1 million myoblasts expressing iRFP and mCardinal or iRFP and mNeptune1. Images were taken in the absence of exogenous biliverdin. The bar graph shows contrast over background (mean ± SEM, n = 6 and n = 8 for mNeptune1 and mCardinal, respectively). Asterisks indicate significant differences by unpaired two-tailed Student t test (* P

Techniques Used: Mouse Assay, Fluorescence, Injection, Expressing, Two Tailed Test

Spectral characteristics of new far-red FPs. (a) Absorbance spectra of oxygenated hemoglobin (oxyHb), deoxygenated hemoglobin (deoxyHb), and monomeric far-red FPs. Myoglobin spectra are similar to hemoglobin spectra 29 . (b) Normalized excitation (left) and emission (right) spectra of monomeric far-red FPs. (c) Transmittance of mKate2, mNeptune1, mNeptune2, mNeptune2.5, mCardinal, and TagRFP657 at 1 mg/mL of purified mature protein. Blue transmission of mCardinal is due to efficient absorbance of green and red light.
Figure Legend Snippet: Spectral characteristics of new far-red FPs. (a) Absorbance spectra of oxygenated hemoglobin (oxyHb), deoxygenated hemoglobin (deoxyHb), and monomeric far-red FPs. Myoglobin spectra are similar to hemoglobin spectra 29 . (b) Normalized excitation (left) and emission (right) spectra of monomeric far-red FPs. (c) Transmittance of mKate2, mNeptune1, mNeptune2, mNeptune2.5, mCardinal, and TagRFP657 at 1 mg/mL of purified mature protein. Blue transmission of mCardinal is due to efficient absorbance of green and red light.

Techniques Used: Purification, Transmission Assay

Non-invasive longitudinal visualization of muscle regeneration in living mice. (a) Tibialis anterior (TA) muscles injected with 1 million myoblasts expressing mCardinal, imaged with a fluorescence stereoscope with 620/20 nm excitation. All images are normalized to the same intensity scale. Series is representative of 5 repeats. (b) Images from 3, 7, and 14 days post-injection (d.p.i.) are shown with intensity scaling tighter than in (a) by a factor of 10, 10, and 5 respectively. (c) Magnified view of the muscle at 7 d.p.i., showing an early regenerating fiber (arrow). The image at right is deliberately enlarged until pixelated to show that this fiber appears just a few pixels wide. (d) Fluorescence signal from TA muscles injected with 1000 muscle stem cells expressing mCardinal. (e) Magnified view of the muscle at 44 d.p.i., showing multiple regenerating fibers (arrows). (f) Bioluminescence imaging at 42 d.p.i. of stem cells shown in (d) . A single 8-min bioluminescence image was acquired at the highest possible resolution with no binning. The resulting sampling resolution (21 μm/pixel) is similar to that (15 μm/pixel) of the fluorescence images in (e). (g) Enlargement of the luminescence image in (f) shows the absence of structures resembling myofibers. The right panel is enlarged to show the presence of noise at the level of individual pixels. In a 40-pixel × 30-pixel region containing the brightest pixels, standard deviation was 33 while mean intensity was 290 counts over that of a background region, indicating presence of substantial shot and read noise relative to signal.
Figure Legend Snippet: Non-invasive longitudinal visualization of muscle regeneration in living mice. (a) Tibialis anterior (TA) muscles injected with 1 million myoblasts expressing mCardinal, imaged with a fluorescence stereoscope with 620/20 nm excitation. All images are normalized to the same intensity scale. Series is representative of 5 repeats. (b) Images from 3, 7, and 14 days post-injection (d.p.i.) are shown with intensity scaling tighter than in (a) by a factor of 10, 10, and 5 respectively. (c) Magnified view of the muscle at 7 d.p.i., showing an early regenerating fiber (arrow). The image at right is deliberately enlarged until pixelated to show that this fiber appears just a few pixels wide. (d) Fluorescence signal from TA muscles injected with 1000 muscle stem cells expressing mCardinal. (e) Magnified view of the muscle at 44 d.p.i., showing multiple regenerating fibers (arrows). (f) Bioluminescence imaging at 42 d.p.i. of stem cells shown in (d) . A single 8-min bioluminescence image was acquired at the highest possible resolution with no binning. The resulting sampling resolution (21 μm/pixel) is similar to that (15 μm/pixel) of the fluorescence images in (e). (g) Enlargement of the luminescence image in (f) shows the absence of structures resembling myofibers. The right panel is enlarged to show the presence of noise at the level of individual pixels. In a 40-pixel × 30-pixel region containing the brightest pixels, standard deviation was 33 while mean intensity was 290 counts over that of a background region, indicating presence of substantial shot and read noise relative to signal.

Techniques Used: Mouse Assay, Injection, Expressing, Fluorescence, Imaging, Sampling, Standard Deviation

9) Product Images from "An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore"

Article Title: An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore

Journal: PLoS ONE

doi: 10.1371/journal.pone.0028674

Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.
Figure Legend Snippet: Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.

Techniques Used: Fluorescence, Imaging, Construct, Expressing

10) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

11) Product Images from "Identification and Functional Comparison of Seven-Transmembrane G-Protein-Coupled BILF1 Receptors in Recently Discovered Nonhuman Primate Lymphocryptoviruses"

Article Title: Identification and Functional Comparison of Seven-Transmembrane G-Protein-Coupled BILF1 Receptors in Recently Discovered Nonhuman Primate Lymphocryptoviruses

Journal: Journal of Virology

doi: 10.1128/JVI.02716-14

Distinct cellular localization patterns of BILF1 receptors. (A) HEK-293 cells were cotransfected with EBV BILF1, PtroLCV1 BILF1, PpygLCV1 BILF1, and SsynLCV1 BILF1 and the farnesylated enhanced green fluorescent protein (eGFPF). The representative pictures
Figure Legend Snippet: Distinct cellular localization patterns of BILF1 receptors. (A) HEK-293 cells were cotransfected with EBV BILF1, PtroLCV1 BILF1, PpygLCV1 BILF1, and SsynLCV1 BILF1 and the farnesylated enhanced green fluorescent protein (eGFPF). The representative pictures

Techniques Used:

12) Product Images from "An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore"

Article Title: An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore

Journal: PLoS ONE

doi: 10.1371/journal.pone.0028674

Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.
Figure Legend Snippet: Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.

Techniques Used: Fluorescence, Imaging, Construct, Expressing

13) Product Images from "A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum"

Article Title: A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum

Journal: Nature methods

doi: 10.1038/nmeth.2413

Fluorescence imaging of mNeonGreen (mNG) fusion vectors. C-terminal mNG fusion constructs (with respect to the fluorescent protein); for each fusion, the linker amino acid length is indicated after the name of the targeted organelle or fusion partner: ( a ) mNG-Annexin (A4)-C-12 (human; plasma membrane); ( b ) mNG-β-actin-C-18 (human; actin cytoskeleton); ( c ) mNG-β-Catenin-C-20 (mouse; tight junctions); ( d ) mNG-CAAX-C-5 (20-amino acid farnesylation signal from c-Ha-Ras; plasma membrane); (e) mNG-CAF1-C-10 (mouse chromatin assembly factor); ( f ) mNG-Caveolin1-C-10 (human); ( g ) mNG-Endosomes-C-14 (human RhoB GTPase); ( h ) mNG-Fascin-C-10 (human; actin bundling); ( i ) mNG-Fibrillarin-C-7 (human; nucleoli); ( j ) mNG-FilaminA-C-14 (human; cytoskeleton); ( k ) mNG-LAMP1-C-20 (rat; lysosomal membrane glycoprotein 1; lysosomes); ( l ) mNG-Clathrin-C-15 (human, light chain B); ( m ) mNG-Myotilin-C-14 (human; actin filaments); ( n ) mNG-PCNA-C-19 (human; replication foci); ( o ) mNG-Plastin-C-10 (human; actin binding); ( p ) mNG-Rab4a-C-7 (human; endosomes); ( q ) mNG-LC3B-C-7 (rat light chain; autophagosoms; ( r ) mNG-Talin-C-18 (mouse; focal adhesions); ( s ) mNG-Tubulin-C-35 (human; microtubules); ( t ) mNG-ZO1-C-14 (human; tight junctions). The cell line used for expression of C-terminal mNG constructs was Madin-Darby canine kidney (MDCK; ATCC, CCL-34) cells in panels ( c ) and ( t ). HeLa CCL2 (ATCC) cells were used in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Fluorescence imaging of mNeonGreen (mNG) fusion vectors. C-terminal mNG fusion constructs (with respect to the fluorescent protein); for each fusion, the linker amino acid length is indicated after the name of the targeted organelle or fusion partner: ( a ) mNG-Annexin (A4)-C-12 (human; plasma membrane); ( b ) mNG-β-actin-C-18 (human; actin cytoskeleton); ( c ) mNG-β-Catenin-C-20 (mouse; tight junctions); ( d ) mNG-CAAX-C-5 (20-amino acid farnesylation signal from c-Ha-Ras; plasma membrane); (e) mNG-CAF1-C-10 (mouse chromatin assembly factor); ( f ) mNG-Caveolin1-C-10 (human); ( g ) mNG-Endosomes-C-14 (human RhoB GTPase); ( h ) mNG-Fascin-C-10 (human; actin bundling); ( i ) mNG-Fibrillarin-C-7 (human; nucleoli); ( j ) mNG-FilaminA-C-14 (human; cytoskeleton); ( k ) mNG-LAMP1-C-20 (rat; lysosomal membrane glycoprotein 1; lysosomes); ( l ) mNG-Clathrin-C-15 (human, light chain B); ( m ) mNG-Myotilin-C-14 (human; actin filaments); ( n ) mNG-PCNA-C-19 (human; replication foci); ( o ) mNG-Plastin-C-10 (human; actin binding); ( p ) mNG-Rab4a-C-7 (human; endosomes); ( q ) mNG-LC3B-C-7 (rat light chain; autophagosoms; ( r ) mNG-Talin-C-18 (mouse; focal adhesions); ( s ) mNG-Tubulin-C-35 (human; microtubules); ( t ) mNG-ZO1-C-14 (human; tight junctions). The cell line used for expression of C-terminal mNG constructs was Madin-Darby canine kidney (MDCK; ATCC, CCL-34) cells in panels ( c ) and ( t ). HeLa CCL2 (ATCC) cells were used in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Fluorescence, Imaging, Construct, Binding Assay, Expressing

14) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

Live cell imaging of mTFP1 fusion vectors . (A)-(D) Laser scanning confocal images of a single HeLa cell expressing mTFP1-H2B-6 (N-terminus; human) progressing through prophase, metaphase, anaphase and interphase, respectively. (E)-(H) Spinning disk confocal images selected from a time-lapse series of HeLa cells expressing mTFP1-annexin (A4)-12 (C-terminus; human) during ionomycin-induced translocation to the plasma and nuclear membranes [35]: (E) 0 min, ionomycin added; (F) 5 min; (G) 7 min; (H) 9 min. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mTFP1 fusion vectors . (A)-(D) Laser scanning confocal images of a single HeLa cell expressing mTFP1-H2B-6 (N-terminus; human) progressing through prophase, metaphase, anaphase and interphase, respectively. (E)-(H) Spinning disk confocal images selected from a time-lapse series of HeLa cells expressing mTFP1-annexin (A4)-12 (C-terminus; human) during ionomycin-induced translocation to the plasma and nuclear membranes [35]: (E) 0 min, ionomycin added; (F) 5 min; (G) 7 min; (H) 9 min. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Expressing, Translocation Assay

15) Product Images from "Voltage-dependent Dynamic FRET Signals from the Transverse Tubules in Mammalian Skeletal Muscle Fibers"

Article Title: Voltage-dependent Dynamic FRET Signals from the Transverse Tubules in Mammalian Skeletal Muscle Fibers

Journal: The Journal of General Physiology

doi: 10.1085/jgp.200709831

Membrane expression of EGFP-F and ECFP-F in FDB muscle fibers. (A) TPLSM image section obtained at the medial plane of a muscle fiber expressing EGFP-F. n denotes nucleus; arrowheads point to the cytoplasmic side of a nucleus; arrows point toward areas of increased fluorescence close to the poles of nuclei. (B) Fluorescence intensity profile obtained from the area delimited by the white rectangle in A. (C and D) TPLSM image sections of an FDB muscle transfected with pECFP-F and stained extracellularly with di-8-ANEPPS. ECFP-F and di-8-ANEPPS fluorescence images are shown in C and D, respectively. (E) Superimposition of image sections in C and D. (F) Normalized fluorescence intensity profiles measured from the area delimited by the rectangles in Fig. 2, C (cyan trace) and D (red trace). The vertical calibration bar is 20 μm and applies to all the images.
Figure Legend Snippet: Membrane expression of EGFP-F and ECFP-F in FDB muscle fibers. (A) TPLSM image section obtained at the medial plane of a muscle fiber expressing EGFP-F. n denotes nucleus; arrowheads point to the cytoplasmic side of a nucleus; arrows point toward areas of increased fluorescence close to the poles of nuclei. (B) Fluorescence intensity profile obtained from the area delimited by the white rectangle in A. (C and D) TPLSM image sections of an FDB muscle transfected with pECFP-F and stained extracellularly with di-8-ANEPPS. ECFP-F and di-8-ANEPPS fluorescence images are shown in C and D, respectively. (E) Superimposition of image sections in C and D. (F) Normalized fluorescence intensity profiles measured from the area delimited by the rectangles in Fig. 2, C (cyan trace) and D (red trace). The vertical calibration bar is 20 μm and applies to all the images.

Techniques Used: Expressing, Fluorescence, Transfection, Staining

Effects of DPA on the electrical and optical properties of a voltage-clamped skeletal muscle fiber expressing EGFP-F. The fiber was clamped at an HP = −90 mV and stimulated with depolarizing and hyperpolarizing voltage pulses of 40, 60, 80, and 100 mV. Data in A and B were obtained before and after DPA staining, respectively. (A) Top panels show capacitive currents in response to 10-ms depolarizing (left) or hyperpolarizing (right) pulses. Bottom panel shows, in an expanded ordinate scale, the corresponding ECFP-F fluorescence records. (B) Capacitive (top) currents and QRET transients (bottom) in response to 20-ms depolarizing (left) and hyperpolarizing (right) voltage-clamp pulses from the same fiber in A, after DPA staining. The rising phase of QRET transients elicited by depolarizing pulses could be adjusted to single exponential functions with τ = 0.4, 1.1, 1.1, and 1.2 ms for the black, red, green, and blue traces, respectively.
Figure Legend Snippet: Effects of DPA on the electrical and optical properties of a voltage-clamped skeletal muscle fiber expressing EGFP-F. The fiber was clamped at an HP = −90 mV and stimulated with depolarizing and hyperpolarizing voltage pulses of 40, 60, 80, and 100 mV. Data in A and B were obtained before and after DPA staining, respectively. (A) Top panels show capacitive currents in response to 10-ms depolarizing (left) or hyperpolarizing (right) pulses. Bottom panel shows, in an expanded ordinate scale, the corresponding ECFP-F fluorescence records. (B) Capacitive (top) currents and QRET transients (bottom) in response to 20-ms depolarizing (left) and hyperpolarizing (right) voltage-clamp pulses from the same fiber in A, after DPA staining. The rising phase of QRET transients elicited by depolarizing pulses could be adjusted to single exponential functions with τ = 0.4, 1.1, 1.1, and 1.2 ms for the black, red, green, and blue traces, respectively.

Techniques Used: Expressing, Staining, Mass Spectrometry, Fluorescence

QRET transient amplitude and DPA//EGFP-F and DPA//ECFP-F spectral overlap. (A) Spectral overlap between DPA absorption spectrum (red trace) and EGFP-F normalized emission spectrum (green trace). (B) Spectral overlap between DPA absorption spectrum (red trace) and ECFP-F normalized emission spectra (cyan trace). EGFP-F and ECFP-F emission spectra were determined in vivo and DPA absorption spectrum was measure in vitro. (C) Peak amplitude of EGFP-F (green hatched bar) and ECFP-F (cyan hatched bar) QRET transients evoked by AP stimulation. The averages values (in −ΔF/F) were calculated from 6 and 12 fibers for EGFP-F and ECFP-F, respectively. The error bar is the SEM. The asterisk indicates statistical significance (P
Figure Legend Snippet: QRET transient amplitude and DPA//EGFP-F and DPA//ECFP-F spectral overlap. (A) Spectral overlap between DPA absorption spectrum (red trace) and EGFP-F normalized emission spectrum (green trace). (B) Spectral overlap between DPA absorption spectrum (red trace) and ECFP-F normalized emission spectra (cyan trace). EGFP-F and ECFP-F emission spectra were determined in vivo and DPA absorption spectrum was measure in vitro. (C) Peak amplitude of EGFP-F (green hatched bar) and ECFP-F (cyan hatched bar) QRET transients evoked by AP stimulation. The averages values (in −ΔF/F) were calculated from 6 and 12 fibers for EGFP-F and ECFP-F, respectively. The error bar is the SEM. The asterisk indicates statistical significance (P

Techniques Used: In Vivo, In Vitro

16) Product Images from "Voltage-dependent Dynamic FRET Signals from the Transverse Tubules in Mammalian Skeletal Muscle Fibers"

Article Title: Voltage-dependent Dynamic FRET Signals from the Transverse Tubules in Mammalian Skeletal Muscle Fibers

Journal: The Journal of General Physiology

doi: 10.1085/jgp.200709831

Membrane expression of EGFP-F and ECFP-F in FDB muscle fibers. (A) TPLSM image section obtained at the medial plane of a muscle fiber expressing EGFP-F. n denotes nucleus; arrowheads point to the cytoplasmic side of a nucleus; arrows point toward areas of increased fluorescence close to the poles of nuclei. (B) Fluorescence intensity profile obtained from the area delimited by the white rectangle in A. (C and D) TPLSM image sections of an FDB muscle transfected with pECFP-F and stained extracellularly with di-8-ANEPPS. ECFP-F and di-8-ANEPPS fluorescence images are shown in C and D, respectively. (E) Superimposition of image sections in C and D. (F) Normalized fluorescence intensity profiles measured from the area delimited by the rectangles in Fig. 2, C (cyan trace) and D (red trace). The vertical calibration bar is 20 μm and applies to all the images.
Figure Legend Snippet: Membrane expression of EGFP-F and ECFP-F in FDB muscle fibers. (A) TPLSM image section obtained at the medial plane of a muscle fiber expressing EGFP-F. n denotes nucleus; arrowheads point to the cytoplasmic side of a nucleus; arrows point toward areas of increased fluorescence close to the poles of nuclei. (B) Fluorescence intensity profile obtained from the area delimited by the white rectangle in A. (C and D) TPLSM image sections of an FDB muscle transfected with pECFP-F and stained extracellularly with di-8-ANEPPS. ECFP-F and di-8-ANEPPS fluorescence images are shown in C and D, respectively. (E) Superimposition of image sections in C and D. (F) Normalized fluorescence intensity profiles measured from the area delimited by the rectangles in Fig. 2, C (cyan trace) and D (red trace). The vertical calibration bar is 20 μm and applies to all the images.

Techniques Used: Expressing, Fluorescence, Transfection, Staining

Effects of DPA on the electrical and optical properties of a voltage-clamped skeletal muscle fiber expressing EGFP-F. The fiber was clamped at an HP = −90 mV and stimulated with depolarizing and hyperpolarizing voltage pulses of 40, 60, 80, and 100 mV. Data in A and B were obtained before and after DPA staining, respectively. (A) Top panels show capacitive currents in response to 10-ms depolarizing (left) or hyperpolarizing (right) pulses. Bottom panel shows, in an expanded ordinate scale, the corresponding ECFP-F fluorescence records. (B) Capacitive (top) currents and QRET transients (bottom) in response to 20-ms depolarizing (left) and hyperpolarizing (right) voltage-clamp pulses from the same fiber in A, after DPA staining. The rising phase of QRET transients elicited by depolarizing pulses could be adjusted to single exponential functions with τ = 0.4, 1.1, 1.1, and 1.2 ms for the black, red, green, and blue traces, respectively.
Figure Legend Snippet: Effects of DPA on the electrical and optical properties of a voltage-clamped skeletal muscle fiber expressing EGFP-F. The fiber was clamped at an HP = −90 mV and stimulated with depolarizing and hyperpolarizing voltage pulses of 40, 60, 80, and 100 mV. Data in A and B were obtained before and after DPA staining, respectively. (A) Top panels show capacitive currents in response to 10-ms depolarizing (left) or hyperpolarizing (right) pulses. Bottom panel shows, in an expanded ordinate scale, the corresponding ECFP-F fluorescence records. (B) Capacitive (top) currents and QRET transients (bottom) in response to 20-ms depolarizing (left) and hyperpolarizing (right) voltage-clamp pulses from the same fiber in A, after DPA staining. The rising phase of QRET transients elicited by depolarizing pulses could be adjusted to single exponential functions with τ = 0.4, 1.1, 1.1, and 1.2 ms for the black, red, green, and blue traces, respectively.

Techniques Used: Expressing, Staining, Mass Spectrometry, Fluorescence

QRET transient amplitude and DPA//EGFP-F and DPA//ECFP-F spectral overlap. (A) Spectral overlap between DPA absorption spectrum (red trace) and EGFP-F normalized emission spectrum (green trace). (B) Spectral overlap between DPA absorption spectrum (red trace) and ECFP-F normalized emission spectra (cyan trace). EGFP-F and ECFP-F emission spectra were determined in vivo and DPA absorption spectrum was measure in vitro. (C) Peak amplitude of EGFP-F (green hatched bar) and ECFP-F (cyan hatched bar) QRET transients evoked by AP stimulation. The averages values (in −ΔF/F) were calculated from 6 and 12 fibers for EGFP-F and ECFP-F, respectively. The error bar is the SEM. The asterisk indicates statistical significance (P
Figure Legend Snippet: QRET transient amplitude and DPA//EGFP-F and DPA//ECFP-F spectral overlap. (A) Spectral overlap between DPA absorption spectrum (red trace) and EGFP-F normalized emission spectrum (green trace). (B) Spectral overlap between DPA absorption spectrum (red trace) and ECFP-F normalized emission spectra (cyan trace). EGFP-F and ECFP-F emission spectra were determined in vivo and DPA absorption spectrum was measure in vitro. (C) Peak amplitude of EGFP-F (green hatched bar) and ECFP-F (cyan hatched bar) QRET transients evoked by AP stimulation. The averages values (in −ΔF/F) were calculated from 6 and 12 fibers for EGFP-F and ECFP-F, respectively. The error bar is the SEM. The asterisk indicates statistical significance (P

Techniques Used: In Vivo, In Vitro

17) Product Images from "Subcellular Localization of the Bovine Leukemia Virus R3 and G4 Accessory Proteins"

Article Title: Subcellular Localization of the Bovine Leukemia Virus R3 and G4 Accessory Proteins

Journal: Journal of Virology

doi: 10.1128/JVI.76.15.7843-7854.2002

R3 is located in cellular membranes and in the nuclear compartment. (A) HeLa Tat cells were cotransfected with pHisR3 and pEGFP-F (Clontech), which contains the 20-amino-acid farnesylation signal from c-Ha-Ras fused to the C terminus of enhanced GFP. Thirty hours posttransfection, cells were fixed, permeabilized, and stained with anti-six-His antibody and Texas Red-conjugated anti-mouse immunoglobulin in order to detect R3. Cells were then analyzed by confocal microscopy. Shown are the signals obtained for GFP-F (green) and R3 (red); the overlay corresponds to the superimposition of the two fluorescent signals. (B) Shown is a quantification of the intensities of both fluorochromes assessed along the line indicated on the overlay in panel A.
Figure Legend Snippet: R3 is located in cellular membranes and in the nuclear compartment. (A) HeLa Tat cells were cotransfected with pHisR3 and pEGFP-F (Clontech), which contains the 20-amino-acid farnesylation signal from c-Ha-Ras fused to the C terminus of enhanced GFP. Thirty hours posttransfection, cells were fixed, permeabilized, and stained with anti-six-His antibody and Texas Red-conjugated anti-mouse immunoglobulin in order to detect R3. Cells were then analyzed by confocal microscopy. Shown are the signals obtained for GFP-F (green) and R3 (red); the overlay corresponds to the superimposition of the two fluorescent signals. (B) Shown is a quantification of the intensities of both fluorochromes assessed along the line indicated on the overlay in panel A.

Techniques Used: Staining, Confocal Microscopy

18) Product Images from "Vinculin is required for cell polarization, migration, and extracellular matrix remodeling in 3D collagen"

Article Title: Vinculin is required for cell polarization, migration, and extracellular matrix remodeling in 3D collagen

Journal: The FASEB Journal

doi: 10.1096/fj.14-268235

Vinculin promotes persistent protrusion and stable orientation of primary MEFs migrating in 3D collagen. A ) Projections of confocal image stacks of live tdTomato-farnesyl–expressing control and Vcl -KO MEFs in 3D collagen; 5 min frame rate. Persistent
Figure Legend Snippet: Vinculin promotes persistent protrusion and stable orientation of primary MEFs migrating in 3D collagen. A ) Projections of confocal image stacks of live tdTomato-farnesyl–expressing control and Vcl -KO MEFs in 3D collagen; 5 min frame rate. Persistent

Techniques Used: Expressing

19) Product Images from "Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging"

Article Title: Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging

Journal: BMC Biology

doi: 10.1186/1741-7007-6-13

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.
Figure Legend Snippet: 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.

Techniques Used: Fluorescence, Imaging, Construct, Sequencing, Expressing

Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Live cell imaging of mWasabi fusion vectors . (A)-(J) 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) mWasabi-α-actinin-19 (human non-muscle); (B) mWasabi-mitochondria-7 (human cytochrome C oxidase subunit VIII); (C) mWasabi-Cx26-7 (rat β-2 connexin-26); (D) mWasabi-keratin-17 (human cytokeratin 18); (E) mWasabi-paxillin-22 (chicken); (F) mWasabi-EB3-7 (human microtubule-associated protein; RP/EB family); (G) mWasabi-golgi-7 (N-terminal 81 amino acids of human β-1,4-galactosyltransferase); (H) mWasabi-vimentin-7 (human); (I)mWasabi-H2B-6 (human); (J) mWasabi-zyxin-7 (human). (K)-(T) C-terminal fusion constructs: (K) mWasabi-lamin B1-10 (human); (L) mWasabi-β-actin-7; (M) mWasabi-β-tubulin-6 (human); (N) mWasabi-clathrin light chain-15 (human); (O) mWasabi-vinculin-23 (human); (P) mWasabi-farnesyl-5 (20-amino acid farnesylation signal from c-Ha-Ras); (Q) mWasabi-focal adhesion kinase-5 (chicken protein tyrosine kinase 2); (R) mWasabi-peroxisomes-2 (peroximal targeting signal 1; PTS1); (S) mWasabi-endosomes-15 (human RhoB GTPase with an N-terminal c-Myc epitope tag); (T) mWasabi-annexin (A4)-15 (human). The cell line used for expressing mWasabi fusion vectors was gray fox lung fibroblast cells (FoLu) in panels (E) and (F) and human cervical adenocarcinoma cells (HeLa) in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Live Cell Imaging, Construct, Expressing

20) Product Images from "An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore"

Article Title: An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore

Journal: PLoS ONE

doi: 10.1371/journal.pone.0028674

Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.
Figure Legend Snippet: Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.

Techniques Used: Fluorescence, Imaging, Construct, Expressing

21) Product Images from "A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum"

Article Title: A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum

Journal: Nature methods

doi: 10.1038/nmeth.2413

Fluorescence imaging of mNeonGreen (mNG) fusion vectors. C-terminal mNG fusion constructs (with respect to the fluorescent protein); for each fusion, the linker amino acid length is indicated after the name of the targeted organelle or fusion partner: ( a ) mNG-Annexin (A4)-C-12 (human; plasma membrane); ( b ) mNG-β-actin-C-18 (human; actin cytoskeleton); ( c ) mNG-β-Catenin-C-20 (mouse; tight junctions); ( d ) mNG-CAAX-C-5 (20-amino acid farnesylation signal from c-Ha-Ras; plasma membrane); (e) mNG-CAF1-C-10 (mouse chromatin assembly factor); ( f ) mNG-Caveolin1-C-10 (human); ( g ) mNG-Endosomes-C-14 (human RhoB GTPase); ( h ) mNG-Fascin-C-10 (human; actin bundling); ( i ) mNG-Fibrillarin-C-7 (human; nucleoli); ( j ) mNG-FilaminA-C-14 (human; cytoskeleton); ( k ) mNG-LAMP1-C-20 (rat; lysosomal membrane glycoprotein 1; lysosomes); ( l ) mNG-Clathrin-C-15 (human, light chain B); ( m ) mNG-Myotilin-C-14 (human; actin filaments); ( n ) mNG-PCNA-C-19 (human; replication foci); ( o ) mNG-Plastin-C-10 (human; actin binding); ( p ) mNG-Rab4a-C-7 (human; endosomes); ( q ) mNG-LC3B-C-7 (rat light chain; autophagosoms; ( r ) mNG-Talin-C-18 (mouse; focal adhesions); ( s ) mNG-Tubulin-C-35 (human; microtubules); ( t ) mNG-ZO1-C-14 (human; tight junctions). The cell line used for expression of C-terminal mNG constructs was Madin-Darby canine kidney (MDCK; ATCC, CCL-34) cells in panels ( c ) and ( t ). HeLa CCL2 (ATCC) cells were used in the remaining panels. Scale bars represent 10 μm.
Figure Legend Snippet: Fluorescence imaging of mNeonGreen (mNG) fusion vectors. C-terminal mNG fusion constructs (with respect to the fluorescent protein); for each fusion, the linker amino acid length is indicated after the name of the targeted organelle or fusion partner: ( a ) mNG-Annexin (A4)-C-12 (human; plasma membrane); ( b ) mNG-β-actin-C-18 (human; actin cytoskeleton); ( c ) mNG-β-Catenin-C-20 (mouse; tight junctions); ( d ) mNG-CAAX-C-5 (20-amino acid farnesylation signal from c-Ha-Ras; plasma membrane); (e) mNG-CAF1-C-10 (mouse chromatin assembly factor); ( f ) mNG-Caveolin1-C-10 (human); ( g ) mNG-Endosomes-C-14 (human RhoB GTPase); ( h ) mNG-Fascin-C-10 (human; actin bundling); ( i ) mNG-Fibrillarin-C-7 (human; nucleoli); ( j ) mNG-FilaminA-C-14 (human; cytoskeleton); ( k ) mNG-LAMP1-C-20 (rat; lysosomal membrane glycoprotein 1; lysosomes); ( l ) mNG-Clathrin-C-15 (human, light chain B); ( m ) mNG-Myotilin-C-14 (human; actin filaments); ( n ) mNG-PCNA-C-19 (human; replication foci); ( o ) mNG-Plastin-C-10 (human; actin binding); ( p ) mNG-Rab4a-C-7 (human; endosomes); ( q ) mNG-LC3B-C-7 (rat light chain; autophagosoms; ( r ) mNG-Talin-C-18 (mouse; focal adhesions); ( s ) mNG-Tubulin-C-35 (human; microtubules); ( t ) mNG-ZO1-C-14 (human; tight junctions). The cell line used for expression of C-terminal mNG constructs was Madin-Darby canine kidney (MDCK; ATCC, CCL-34) cells in panels ( c ) and ( t ). HeLa CCL2 (ATCC) cells were used in the remaining panels. Scale bars represent 10 μm.

Techniques Used: Fluorescence, Imaging, Construct, Binding Assay, Expressing

22) Product Images from "Subcellular Localization of the Bovine Leukemia Virus R3 and G4 Accessory Proteins"

Article Title: Subcellular Localization of the Bovine Leukemia Virus R3 and G4 Accessory Proteins

Journal: Journal of Virology

doi: 10.1128/JVI.76.15.7843-7854.2002

R3 is located in cellular membranes and in the nuclear compartment. (A) HeLa Tat cells were cotransfected with pHisR3 and pEGFP-F (Clontech), which contains the 20-amino-acid farnesylation signal from c-Ha-Ras fused to the C terminus of enhanced GFP. Thirty hours posttransfection, cells were fixed, permeabilized, and stained with anti-six-His antibody and Texas Red-conjugated anti-mouse immunoglobulin in order to detect R3. Cells were then analyzed by confocal microscopy. Shown are the signals obtained for GFP-F (green) and R3 (red); the overlay corresponds to the superimposition of the two fluorescent signals. (B) Shown is a quantification of the intensities of both fluorochromes assessed along the line indicated on the overlay in panel A.
Figure Legend Snippet: R3 is located in cellular membranes and in the nuclear compartment. (A) HeLa Tat cells were cotransfected with pHisR3 and pEGFP-F (Clontech), which contains the 20-amino-acid farnesylation signal from c-Ha-Ras fused to the C terminus of enhanced GFP. Thirty hours posttransfection, cells were fixed, permeabilized, and stained with anti-six-His antibody and Texas Red-conjugated anti-mouse immunoglobulin in order to detect R3. Cells were then analyzed by confocal microscopy. Shown are the signals obtained for GFP-F (green) and R3 (red); the overlay corresponds to the superimposition of the two fluorescent signals. (B) Shown is a quantification of the intensities of both fluorochromes assessed along the line indicated on the overlay in panel A.

Techniques Used: Staining, Confocal Microscopy

) into plasmids pEGFP-C1 (Clontech) and pcDNA3.1/HisB (Invitrogen), respectively. To generate pR3GFP, R3 was PCR amplified with primers Hin dIIIS (5′-TTTAAGCTTGGCTTTAAAATGGCTAAAGAACG-3′) and TAAR3Bam (5′-TTTGGATCCGAGAAAAGGCGGCCCAAATGC-3′), digested with Hin dIII/ Bam HI, and inserted in frame upstream of the enhanced GFP gene in plasmid pEGFP-N1 (Clontech). HeLa Tat cells were transfected with pHisR3 (A), pEGFPR3 (B), or pR3GFP (C). Thirty hours posttransfection, cells were fixed and examined directly by confocal microscopy (B and C) or fixed, permeabilized, and incubated with a monoclonal anti-six-His antibody (Sigma) and FITC-coupled anti-mouse immunoglobulin (A).
Figure Legend Snippet: ) into plasmids pEGFP-C1 (Clontech) and pcDNA3.1/HisB (Invitrogen), respectively. To generate pR3GFP, R3 was PCR amplified with primers Hin dIIIS (5′-TTTAAGCTTGGCTTTAAAATGGCTAAAGAACG-3′) and TAAR3Bam (5′-TTTGGATCCGAGAAAAGGCGGCCCAAATGC-3′), digested with Hin dIII/ Bam HI, and inserted in frame upstream of the enhanced GFP gene in plasmid pEGFP-N1 (Clontech). HeLa Tat cells were transfected with pHisR3 (A), pEGFPR3 (B), or pR3GFP (C). Thirty hours posttransfection, cells were fixed and examined directly by confocal microscopy (B and C) or fixed, permeabilized, and incubated with a monoclonal anti-six-His antibody (Sigma) and FITC-coupled anti-mouse immunoglobulin (A).

Techniques Used: Polymerase Chain Reaction, Amplification, Plasmid Preparation, Transfection, Confocal Microscopy, Incubation

23) Product Images from "An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore"

Article Title: An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore

Journal: PLoS ONE

doi: 10.1371/journal.pone.0028674

Fluorescence imaging of mTagBFP2 fusion constructs. (A–M)  C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X)  N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.
Figure Legend Snippet: Fluorescence imaging of mTagBFP2 fusion constructs. (A–M) C-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) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.

Techniques Used: Fluorescence, Imaging, Construct, Expressing

Related Articles

Clone Assay:

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Article Snippet: .. Cloning The region corresponding to 2B was amplified from the cDNA of HAV strain HM175, and cloned into the NheI and BamHI sites of the expression vector pEGFP-N1 (Clonetech). .. The point mutation V216A, to convert the sequence into wildtype from a cell culture adapted strain, was generated by site-directed mutagenesis (Stratagene).

Article Title: Mutations in QARS, Encoding Glutaminyl-tRNA Synthetase, Cause Progressive Microcephaly, Cerebral-Cerebellar Atrophy, and Intractable Seizures
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shRNA:

Article Title: Expression level is a key determinant of E2F1-mediated cell fate
Article Snippet: .. pEYFP-ER-E2F1 construct was generated by inserting the Nhe I digestion fragment containing the ER-E2F1 cassette from pBabePuro-HA.ER.E2F1 (a gift from Dr. Kristian Helin) into the compatible Xba I digestion site of the pEYFP-C1 vector (Clontech, Mountain View, CA, USA). pTRIPZ-YFP-ER-E2F1 construct was generated by inserting the YFP-ER-E2F1 cassette from pEYFP-ER-E2F1 into the pTRIPZ vector (Open Biosystems, Lafayette, CO, USA) between the Age I and Mlu I sites replacing the TurboRFP and shRNA cassette. .. For CRISPR-Cas9-mediated E2F1 knockout, gRNA sequence 5′-GGAGATGATGACGATCTGCG-3′ targeting exon 1 of E2F1 was cloned into LentiCRISPR v.2 (Addgene, Cambridge, MA, USA, #52961).

Incubation:

Article Title: The S-phase Cyclin Clb5 Promotes rDNA Stability by Maintaining Replication Initiation Efficiency in rDNA
Article Snippet: .. After discarding the buffer completely, the plug was incubated in 160 μl of 1× M buffer containing 160 units of Nhe I (TaKaRa) for 7 hr at 37°C. .. The plug and 600 ng of Lambda Hind III DNA markers were separated by electrophoresis on a 0.4% agarose gel (SeaKem Agarose LE, Lonza) in 1× TBE buffer at 1.32 V/cm for 14 hr at room temperature with buffer circulation in a Sub-cell GT electrophoresis system (15 × 20 cm gel, Bio-Rad).

Construct:

Article Title: Expression level is a key determinant of E2F1-mediated cell fate
Article Snippet: .. pEYFP-ER-E2F1 construct was generated by inserting the Nhe I digestion fragment containing the ER-E2F1 cassette from pBabePuro-HA.ER.E2F1 (a gift from Dr. Kristian Helin) into the compatible Xba I digestion site of the pEYFP-C1 vector (Clontech, Mountain View, CA, USA). pTRIPZ-YFP-ER-E2F1 construct was generated by inserting the YFP-ER-E2F1 cassette from pEYFP-ER-E2F1 into the pTRIPZ vector (Open Biosystems, Lafayette, CO, USA) between the Age I and Mlu I sites replacing the TurboRFP and shRNA cassette. .. For CRISPR-Cas9-mediated E2F1 knockout, gRNA sequence 5′-GGAGATGATGACGATCTGCG-3′ targeting exon 1 of E2F1 was cloned into LentiCRISPR v.2 (Addgene, Cambridge, MA, USA, #52961).

Produced:

Article Title: Good Manufacturing Practices production and analysis of a DNA vaccine against dental caries
Article Snippet: .. Single identification digestion was made using only one restriction enzyme, Kpn I (Takara Bio Inc, Otsu, Japan) or Nhe I (Takara Bio Inc, Otsu, Japan), both of which produced one fragment 7349 bp in size; or Xho I (Takara Bio Inc, Otsu, Japan), which produced two fragments of 2273 bp and 5076 bp. ..

Article Title: Good Manufacturing Practices production and analysis of a DNA vaccine against dental caries
Article Snippet: .. Single identification digestion was made using only one restriction enzyme, Kpn I (Takara Bio Inc, Otsu, Japan) or Nhe I (Takara Bio Inc, Otsu, Japan), both of which produced one fragment 7349 bp in size; or Xho I (Takara Bio Inc, Otsu, Japan), which produced two fragments of 2273 bp and 5076 bp. ..

Generated:

Article Title: Expression level is a key determinant of E2F1-mediated cell fate
Article Snippet: .. pEYFP-ER-E2F1 construct was generated by inserting the Nhe I digestion fragment containing the ER-E2F1 cassette from pBabePuro-HA.ER.E2F1 (a gift from Dr. Kristian Helin) into the compatible Xba I digestion site of the pEYFP-C1 vector (Clontech, Mountain View, CA, USA). pTRIPZ-YFP-ER-E2F1 construct was generated by inserting the YFP-ER-E2F1 cassette from pEYFP-ER-E2F1 into the pTRIPZ vector (Open Biosystems, Lafayette, CO, USA) between the Age I and Mlu I sites replacing the TurboRFP and shRNA cassette. .. For CRISPR-Cas9-mediated E2F1 knockout, gRNA sequence 5′-GGAGATGATGACGATCTGCG-3′ targeting exon 1 of E2F1 was cloned into LentiCRISPR v.2 (Addgene, Cambridge, MA, USA, #52961).

Amplification:

Article Title: The C-terminal region of the non-structural protein 2B from Hepatitis A Virus demonstrates lipid-specific viroporin-like activity
Article Snippet: .. Cloning The region corresponding to 2B was amplified from the cDNA of HAV strain HM175, and cloned into the NheI and BamHI sites of the expression vector pEGFP-N1 (Clonetech). .. The point mutation V216A, to convert the sequence into wildtype from a cell culture adapted strain, was generated by site-directed mutagenesis (Stratagene).

Expressing:

Article Title: The C-terminal region of the non-structural protein 2B from Hepatitis A Virus demonstrates lipid-specific viroporin-like activity
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Polymerase Chain Reaction:

Article Title: The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells
Article Snippet: .. The PCR products were recovered and their sequences were confirmed, followed by digestion with Ase I and Nhe I (Takara Biotechnology Co., Ltd., Dalian, China). .. The products were then ligated into the pEM vector to obtain vectors containing the EF‐1α or CAG promoter (Fig. D and E).

Article Title: Receptor-interacting protein 140 is a repressor of the androgen receptor activity
Article Snippet: .. pCMV-AR and pSG5-TIF2 were generous gifts from respectively Drs Terry Brown and Hinrich Gronemeyer. pCMV-AR(1-660), pCMV-AR(507-919) ( ) and pFC31-Luc ( ) were already described. pEYFP-RIP140, pcRIP140, pcRIP(1-282), pcRIP(1-480), pcRIP(479-1158), pcRIP(917-1158), pcRIPmutPIDLS, pcRIPmutPINLS and pcRIPmutPID/NLS were described elsewhere ( ). pEF-c-mycRIP140 was created by inserting the PCR-amplified cDNA encoding c-myc fused to RIP140 amino-acids 1 to 479 into pEF-RIP140 previously digested with BclI and EcoRV. pEF-RIPmutPIDLS was created by digestion of pcRIPmutPIDLS with BclI and EcoRV and insertion of the resulting fragment into pEF-RIP140. pEF-RIPmutPINLS was created by digestion of pcRIPmutPINLS with EcoRV and BlpI and insertion of the resulting fragment into pEF-RIP140. pEF-RIPmutPID/NLS was created by digestion of pcRIPmutPID/NLS with BclI and BlpI and insertion of the resulting fragment into pEF-RIP140. pECFP-AR(507-919) was created by inserting the PCR-amplified cDNA encoding AR(507-919) into pECFP-C2 previously digested with BamHI. pGFP-AR(1-501) was digested with XhoI and XbaI and the resulting fragment was inserted into pECFP-AR previously digested with the same enzymes to create pECFP-AR(1-501). pECFP-AR was obtained by insertion of AR excised by NheI and BglII restriction sites from the previously described pGFP-AR ( ) into pECFP-C1 (Clontech, Palo Alto, CA). ..

Article Title: Mutations in QARS, Encoding Glutaminyl-tRNA Synthetase, Cause Progressive Microcephaly, Cerebral-Cerebellar Atrophy, and Intractable Seizures
Article Snippet: .. Human QARS was codon optimized for E. coli strains (GeneArt) and cloned between NheI and XhoI sites of pET28a for the generation of an N-terminal His-tagged fusion protein with the use of the In-Fusion PCR Cloning System (Clontech). .. Point mutations c.134G > T, c.1207C > T, c.169T > C, and c.1543C > T were introduced with the QuickChange mutagenesis method. pET28a plasmids with WT QARS and mutant alleles were transformed into the Rosetta pLysS strain.

Plasmid Preparation:

Article Title: Expression level is a key determinant of E2F1-mediated cell fate
Article Snippet: .. pEYFP-ER-E2F1 construct was generated by inserting the Nhe I digestion fragment containing the ER-E2F1 cassette from pBabePuro-HA.ER.E2F1 (a gift from Dr. Kristian Helin) into the compatible Xba I digestion site of the pEYFP-C1 vector (Clontech, Mountain View, CA, USA). pTRIPZ-YFP-ER-E2F1 construct was generated by inserting the YFP-ER-E2F1 cassette from pEYFP-ER-E2F1 into the pTRIPZ vector (Open Biosystems, Lafayette, CO, USA) between the Age I and Mlu I sites replacing the TurboRFP and shRNA cassette. .. For CRISPR-Cas9-mediated E2F1 knockout, gRNA sequence 5′-GGAGATGATGACGATCTGCG-3′ targeting exon 1 of E2F1 was cloned into LentiCRISPR v.2 (Addgene, Cambridge, MA, USA, #52961).

Article Title: The C-terminal region of the non-structural protein 2B from Hepatitis A Virus demonstrates lipid-specific viroporin-like activity
Article Snippet: .. Cloning The region corresponding to 2B was amplified from the cDNA of HAV strain HM175, and cloned into the NheI and BamHI sites of the expression vector pEGFP-N1 (Clonetech). .. The point mutation V216A, to convert the sequence into wildtype from a cell culture adapted strain, was generated by site-directed mutagenesis (Stratagene).

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