imaris colocalization analysis Search Results


colocalization analysis  (Oxford Instruments)
99
Oxford Instruments colocalization analysis
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Colocalization Analysis, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/Imaris/pmc02064081-251-21-26
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nis elements software  (Nikon)
99
Nikon nis elements software
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Nis Elements Software, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/NIS-Elements/pmc07227422-229-10-9
Average 99 stars, based on 1 article reviews
nis elements software - by Bioz Stars, 2026-10
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vectra polaris microscope  (Vectra Laboratories)
86
Vectra Laboratories vectra polaris microscope
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Vectra Polaris Microscope, supplied by Vectra Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/polaris+vectra/pm42135303-927-4-4
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imspector image acquisition & analysis software v16.3  (abberior instruments)
90
abberior instruments imspector image acquisition & analysis software v16.3
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Imspector Image Acquisition & Analysis Software V16.3, supplied by abberior instruments, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/imspector+image+acquisition+++analysis+software+v16+3/pm37948187-181-152-141
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nbp2 22399 anti lamp1 rabbit  (Rockland Immunochemicals)
93
Rockland Immunochemicals nbp2 22399 anti lamp1 rabbit
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Nbp2 22399 Anti Lamp1 Rabbit, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/LAMP1+Antibody/pm37948187-181-12-23
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cellulose-bound peptide arrays - celluspots  (Proteomic Solutions)
90
Proteomic Solutions cellulose-bound peptide arrays - celluspots
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Cellulose Bound Peptide Arrays Celluspots, supplied by Proteomic Solutions, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/cellulose+bound+peptide+arrays+++celluspots/pm33979625-251-204-205
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fluorsavetm reagent  (Merck KGaA)
90
Merck KGaA fluorsavetm reagent
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Fluorsavetm Reagent, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/fluorsavetm+reagent/pm33979625-251-186-188
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phosphatase inhibitor cocktail  (Merck KGaA)
90
Merck KGaA phosphatase inhibitor cocktail
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Phosphatase Inhibitor Cocktail, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/phosphatase+inhibitor+cocktail/pm33979625-251-129-133
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1003 plasmacytoid dendritic cell kit ii negative purification miltenyi biotec 130 097 415  (Miltenyi Biotec)
95
Miltenyi Biotec 1003 plasmacytoid dendritic cell kit ii negative purification miltenyi biotec 130 097 415
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
1003 Plasmacytoid Dendritic Cell Kit Ii Negative Purification Miltenyi Biotec 130 097 415, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/Plasmacytoid+Dendritic+Cell+Isolation+Kit+II%2C+human/pmc05640662__41467_2017_695_MOESM1_ESM-119-48-56
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1003 plasmacytoid dendritic cell kit ii negative purification miltenyi biotec 130 097 415 - by Bioz Stars, 2026-10
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criterion xt bis tris protein gel bio rad  (Bio-Rad)
96
Bio-Rad criterion xt bis tris protein gel bio rad
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Criterion Xt Bis Tris Protein Gel Bio Rad, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/imaris+colocalization+analysis/4%E2%80%9312%25+Criterion+XT+Bis-Tris+Protein+Gel/pm33979625-251-94-99
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recombinant dna reagent mouse pgap2 plasmid origene origene  (OriGene)
90
OriGene recombinant dna reagent mouse pgap2 plasmid origene origene
MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and <t>colocalization</t> in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).
Recombinant Dna Reagent Mouse Pgap2 Plasmid Origene Origene, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and colocalization in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).

Journal: The Journal of Cell Biology

Article Title: Missing-in-metastasis and IRSp53 deform PI(4,5)P 2 -rich membranes by an inverse BAR domain–like mechanism

doi: 10.1083/jcb.200609176

Figure Lengend Snippet: MIM/IMD-L localizes to the plasma membrane in filopodia and associates actin to membranes in vitro. (A) GFP–MIM/IMD-L localized to the plasma membrane and coated the F-actin bundles in filopodia of U2OS cell. 3D analysis derived from seven confocal planes shows F-actin in red (all panels), MIM/IMD in green (right), and colocalization in yellow (middle and right). Colocalization area (built by ImarisColoc software) is only a thin layer between actin and MIM/IMD and is almost invisible when the three channels are merged (right), as the MIM/IMD signal at the plasma membrane covers the interior of filopodia. Bar, 1 μm. (B) In vitro light microscopy assay. 1 μM F-actin (50% Alexa 568–labeled), 2.5 μM MIM/IMD-L, and 1.67 μM lipid vesicles (0/30% PI[4,5]P 2 ) were mixed and applied on polyornithine-coated glass slides for imaging. In the presence of MIM/IMD-L and PI(4,5)P 2 , actin concentrated at the sites of vesicles, indicating MIM/IMD-L–associating actin and vesicles (top). MIM/IMD-L with vesicles without PI(4,5)P 2 or MIM/IMD-L Mut15, which displays defects in actin and PI(4,5)P 2 binding, induced much weaker colocalization between actin and vesicles. Red arrowheads indicate vesicles that show clear colocalization with actin, and green arrowheads indicate vesicles without colocalizing actin. Bar, 20 μm. (C) The intensity of the actin staining on the vesicles was quantified from 19–20 randomly selected vesicles. Error bars indicate SEM values. A t test was used to analyze p-values and showed statistically significant differences (***, P < 0.001; **, P < 0.01).

Article Snippet: The confocal image stacks were deconvoluted with AutoQuant AutoDeblur 3D Blind Deconvolution (AutoQuant Imaging, Inc.), and the 3D reconstructions and the colocalization analysis were made with Bitplane Imaris (Bitplane Inc.).

Techniques: Membrane, In Vitro, Derivative Assay, Software, Light Microscopy, Labeling, Imaging, Binding Assay, Staining