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  • 99
    Carl Zeiss microscope
    Effects of evolved mutations in the pbpA operon on cell morphology. Cells were grown in DM250 with or without amdinocillin (Mecillinam) and observed using a Zeiss <t>Axioplan</t> 2 microscope. All photographs were obtained using a magnification of ×1,000,
    Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 99/100, based on 60569 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Carl Zeiss confocal microscope
    Effects of evolved mutations in the pbpA operon on cell morphology. Cells were grown in DM250 with or without amdinocillin (Mecillinam) and observed using a Zeiss <t>Axioplan</t> 2 microscope. All photographs were obtained using a magnification of ×1,000,
    Confocal Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 99/100, based on 47725 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Carl Zeiss confocal laser scanning microscope
    Localization of overexpressed torsinA proteins in hippocampal neurons of rats and ΔE-torsinA knock-in mice. Cultured neurons were transfected with a construct encoding either WT-torsinA-GFP ( A ) or ΔE-torsinA-GFP ( B ). A modification of the calcium-phosphate method in Fig. 1 was used to enhance the efficiency of transfection ( Jiang and Chen, 2006 ). A <t>confocal</t> <t>laser-scanning</t> <t>microscope</t> was used to observe the neurons. Neurons are shown both as single optical sections at the somatic level (Single) and as maximum intensity projection (MIP) images that represent the overall structure at all heights. Neurons were cultured from WT rats (top row), WT mice (2 nd row), heterozygous (HET, Tor1a +/ΔE , 3 rd row) or homozygous (HOM, Tor1a ΔE/ΔE , bottom row) ΔE-torsinA knock-in mice. Arrows point to representative examples of the diffuse distribution of WT-torsinA-GFP in the cytoplasm ( A ), and ΔE-torsinA-GFP in cytoplasmic inclusion bodies ( B ).
    Confocal Laser Scanning Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 95/100, based on 39975 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Olympus olympus bx51 microscope
    Anti-Neu humoral response following rV- neu T vaccination. Panel A : p185 Neu expression in NIH3T3 cells encoding normal rat Neu (LTR-Neu) and in Neu-overexpressing BALB- neu T salivary gland cancer (SALTO) cells by western blotting. A specific polyclonal anti-Neu (C18) antibody was used. NIH3T3 cells were used as negative control. Panel B : Igs from rV- neu T vaccinated mice recognize p185 Neu expressed on the surface of SALTO tumor cells. Immunofluorescence was performed using purified Igs (1μg/ml) of BALB- neu T mice vaccinated with rV- neu T or V-wt. The specific monoclonal antibody anti-Neu Ab4 was used as positive control. Olympus <t>BX51</t> microscope and I.A.S. version 007 000 Deconvolution 2D software were used to deconvolve z series images of stained native cells. Original magnification x400. Panel C : Serum antibody response of mice upon vaccination with rV- neu T. Numbering identifies immune sera of individual mice. Mouse pre-immune or immune sera were collected prior to vaccination or one week after the second immunization and employed in immunoprecipitation of Neu from LTR-Neu or SALTO tumor cells. p185 Neu specificity was visualized by immunoblotting analysis using receptor-specific polyclonal antibody of immunoprecipitates and compared to direct immunoblotting of LTR-Neu lysate as positive control, or NIH3T3 as negative control.
    Olympus Bx51 Microscope, supplied by Olympus, used in various techniques. Bioz Stars score: 97/100, based on 30804 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Olympus confocal microscope
    Anti-Neu humoral response following rV- neu T vaccination. Panel A : p185 Neu expression in NIH3T3 cells encoding normal rat Neu (LTR-Neu) and in Neu-overexpressing BALB- neu T salivary gland cancer (SALTO) cells by western blotting. A specific polyclonal anti-Neu (C18) antibody was used. NIH3T3 cells were used as negative control. Panel B : Igs from rV- neu T vaccinated mice recognize p185 Neu expressed on the surface of SALTO tumor cells. Immunofluorescence was performed using purified Igs (1μg/ml) of BALB- neu T mice vaccinated with rV- neu T or V-wt. The specific monoclonal antibody anti-Neu Ab4 was used as positive control. Olympus <t>BX51</t> microscope and I.A.S. version 007 000 Deconvolution 2D software were used to deconvolve z series images of stained native cells. Original magnification x400. Panel C : Serum antibody response of mice upon vaccination with rV- neu T. Numbering identifies immune sera of individual mice. Mouse pre-immune or immune sera were collected prior to vaccination or one week after the second immunization and employed in immunoprecipitation of Neu from LTR-Neu or SALTO tumor cells. p185 Neu specificity was visualized by immunoblotting analysis using receptor-specific polyclonal antibody of immunoprecipitates and compared to direct immunoblotting of LTR-Neu lysate as positive control, or NIH3T3 as negative control.
    Confocal Microscope, supplied by Olympus, used in various techniques. Bioz Stars score: 96/100, based on 21812 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Carl Zeiss lsm 510 confocal microscope
    Heterozygosity for Cldn14 increases tumour blood vessel leakage and decreases intratumoural hypoxia. Wild-type, Cldn14-heterozygous and Cldn14-null mice were injected subcutaneously in the flank with 0.5×10 6 B16F10 melanoma or Lewis Lung Carcinoma (LLC) cells. ( A ) At 10 days post inoculation, PE-conjugated anti-PECAM antibody and Hoechst dye were injected via the tail vein prior to sacrifice. Midline sections (100 µm) of snap-frozen tumours were fixed, mounted and imaged using a Zeiss <t>LSM</t> 510 confocal microscope. The extent of Hoechst leakage was measured in z-stacks using ImageJ. Bars show mean Hoechst leakage relative to PECAM signal ± SEM. Blood vessel leakage is increased significantly in Cldn14-het mice when compared with WT and Cldn14-null mice. ( B ) Representative images of Hoechst (blue) and PECAM (red) detection. ( C ) Tumour-bearing mice from each genotype were injected with pimonidazole prior to sacrifice to measure hypoxic areas within the tumour. 8 µm tumour cryosections were then double stained with anti-pimonidazole antibody (green) to highlight hypoxic areas and anti-PECAM antibody to identify blood vessels. The hypoxic index was quantified relative to PECAM staining using image J software. Bars represent mean relative hypoxic index ± SEM. ( D ) Representative images of pimonidazole detection and PECAM-positive blood vessels in tumour sections. Arrows , blood vessels; Asterisks , pimonidazole-positive staining. Scale bars: A 50 µm; D 200 µm. N = 4 tumours per genotype. NSD: not statistically different, * P
    Lsm 510 Confocal Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 96/100, based on 25422 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    JEOL transmission electron microscope
    Heterozygosity for Cldn14 increases tumour blood vessel leakage and decreases intratumoural hypoxia. Wild-type, Cldn14-heterozygous and Cldn14-null mice were injected subcutaneously in the flank with 0.5×10 6 B16F10 melanoma or Lewis Lung Carcinoma (LLC) cells. ( A ) At 10 days post inoculation, PE-conjugated anti-PECAM antibody and Hoechst dye were injected via the tail vein prior to sacrifice. Midline sections (100 µm) of snap-frozen tumours were fixed, mounted and imaged using a Zeiss <t>LSM</t> 510 confocal microscope. The extent of Hoechst leakage was measured in z-stacks using ImageJ. Bars show mean Hoechst leakage relative to PECAM signal ± SEM. Blood vessel leakage is increased significantly in Cldn14-het mice when compared with WT and Cldn14-null mice. ( B ) Representative images of Hoechst (blue) and PECAM (red) detection. ( C ) Tumour-bearing mice from each genotype were injected with pimonidazole prior to sacrifice to measure hypoxic areas within the tumour. 8 µm tumour cryosections were then double stained with anti-pimonidazole antibody (green) to highlight hypoxic areas and anti-PECAM antibody to identify blood vessels. The hypoxic index was quantified relative to PECAM staining using image J software. Bars represent mean relative hypoxic index ± SEM. ( D ) Representative images of pimonidazole detection and PECAM-positive blood vessels in tumour sections. Arrows , blood vessels; Asterisks , pimonidazole-positive staining. Scale bars: A 50 µm; D 200 µm. N = 4 tumours per genotype. NSD: not statistically different, * P
    Transmission Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 96/100, based on 20889 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Nikon light microscope
    Heterozygosity for Cldn14 increases tumour blood vessel leakage and decreases intratumoural hypoxia. Wild-type, Cldn14-heterozygous and Cldn14-null mice were injected subcutaneously in the flank with 0.5×10 6 B16F10 melanoma or Lewis Lung Carcinoma (LLC) cells. ( A ) At 10 days post inoculation, PE-conjugated anti-PECAM antibody and Hoechst dye were injected via the tail vein prior to sacrifice. Midline sections (100 µm) of snap-frozen tumours were fixed, mounted and imaged using a Zeiss <t>LSM</t> 510 confocal microscope. The extent of Hoechst leakage was measured in z-stacks using ImageJ. Bars show mean Hoechst leakage relative to PECAM signal ± SEM. Blood vessel leakage is increased significantly in Cldn14-het mice when compared with WT and Cldn14-null mice. ( B ) Representative images of Hoechst (blue) and PECAM (red) detection. ( C ) Tumour-bearing mice from each genotype were injected with pimonidazole prior to sacrifice to measure hypoxic areas within the tumour. 8 µm tumour cryosections were then double stained with anti-pimonidazole antibody (green) to highlight hypoxic areas and anti-PECAM antibody to identify blood vessels. The hypoxic index was quantified relative to PECAM staining using image J software. Bars represent mean relative hypoxic index ± SEM. ( D ) Representative images of pimonidazole detection and PECAM-positive blood vessels in tumour sections. Arrows , blood vessels; Asterisks , pimonidazole-positive staining. Scale bars: A 50 µm; D 200 µm. N = 4 tumours per genotype. NSD: not statistically different, * P
    Light Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 16885 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Carl Zeiss zeiss microscope
    PDGF-AA-induced dorsal ruffle formation is diminished in WIP −/− fibroblasts. a Control (WIP +/+ ) and WIP −/− primary murine fibroblasts were serum starved over night (0 min) or serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and <t>FITC-secondary</t> antibody and imaged in a <t>Zeiss</t> microscope to identify dorsal ruffles (white arrow). Magnifications of the boxed areas are shown as right panels. b The percentage of WIP +/+ (black) and WIP −/− (white) cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. c WIP −/− primary fibroblasts were lentivirally transduced to express control cherry or WIP-cherry, starved and incubated with PDGF-AA for 8 or 15 min. Fixed cells were imaged. d The percentage of WIP −/− cells expressing cherry (white) or WIP-cherry (black) and forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. e Representative western blot of PDGFRα expression in soluble lysates of lung-derived fibroblasts from WIP +/+ and WIP −/− mice. Numbers indicate relative expression levels of the protein to control fibroblasts determined by densitometry. ß-actin labeling confirmed equivalent protein loading control. f WIP +/+ (black) and WIP −/− (white) primary fibroblasts were grown in the presence of serum (left panel) and stained with anti-PDGFRα plus labeled secondary antibody and analysed by FACS. The mean fluorescence intensity of positive cells in both populations is represented. Right panel, WIP +/+ (black) and WIP −/− (white) primary fibroblasts were starved (time 0) and stimulated with PDGF-AA and stained as above. Percent of mean fluorescence intensity relative to starvation is represented. Arbitrary units (a.u.). * p
    Zeiss Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 99/100, based on 17513 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    JEOL scanning electron microscope
    PDGF-AA-induced dorsal ruffle formation is diminished in WIP −/− fibroblasts. a Control (WIP +/+ ) and WIP −/− primary murine fibroblasts were serum starved over night (0 min) or serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and <t>FITC-secondary</t> antibody and imaged in a <t>Zeiss</t> microscope to identify dorsal ruffles (white arrow). Magnifications of the boxed areas are shown as right panels. b The percentage of WIP +/+ (black) and WIP −/− (white) cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. c WIP −/− primary fibroblasts were lentivirally transduced to express control cherry or WIP-cherry, starved and incubated with PDGF-AA for 8 or 15 min. Fixed cells were imaged. d The percentage of WIP −/− cells expressing cherry (white) or WIP-cherry (black) and forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. e Representative western blot of PDGFRα expression in soluble lysates of lung-derived fibroblasts from WIP +/+ and WIP −/− mice. Numbers indicate relative expression levels of the protein to control fibroblasts determined by densitometry. ß-actin labeling confirmed equivalent protein loading control. f WIP +/+ (black) and WIP −/− (white) primary fibroblasts were grown in the presence of serum (left panel) and stained with anti-PDGFRα plus labeled secondary antibody and analysed by FACS. The mean fluorescence intensity of positive cells in both populations is represented. Right panel, WIP +/+ (black) and WIP −/− (white) primary fibroblasts were starved (time 0) and stimulated with PDGF-AA and stained as above. Percent of mean fluorescence intensity relative to starvation is represented. Arbitrary units (a.u.). * p
    Scanning Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 95/100, based on 10036 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Olympus confocal laser scanning microscope
    Localization of Mel-AF on A375 cells was analyzed using a <t>confocal</t> <t>laser</t> <t>scanning</t> <t>microscope.</t> The cells were treated with FITC-conjugated Mel-AF for 2 h. ( A ) Confocal images show the localization of Mel-AF traced with FITC (green). The membrane compartment was indicated by staining with a specific antibody to CD46, a membrane cofactor protein followed by Alexa Fluor 568 rabbit anti-mouse IgG (red). The cellular nuclei were counterstained by DAPI (blue). ( B ) Orthogonal imaging analysis was performed to confirm the localization of Mel-AF on the cell membrane (yellow). Scale bar, 10 μm.
    Confocal Laser Scanning Microscope, supplied by Olympus, used in various techniques. Bioz Stars score: 96/100, based on 18562 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Carl Zeiss lsm 710 confocal microscope
    Optochemical activation of Eg5 siRNA in HeLa cells. HeLa cells were transfected with caged and non-caged siRNAs (40 pmol). The cells were irradiated (5 min, 25 W, 365 nm) and incubated at 37°C, 5% CO 2 for 48 h. ( A–G ) The cells were fixed and stained with Alexa Fluor 488 phalloidin (green) and DAPI (blue). The cells were imaged on a Zeiss <t>LSM</t> 710 confocal microscope using a 40× oil objective and Alexa Fluor 488 and DAPI-specific lasers (488 nm multiline argon and 405 nm diode).
    Lsm 710 Confocal Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 96/100, based on 16007 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Hitachi Ltd scanning electron microscope
    Optochemical activation of Eg5 siRNA in HeLa cells. HeLa cells were transfected with caged and non-caged siRNAs (40 pmol). The cells were irradiated (5 min, 25 W, 365 nm) and incubated at 37°C, 5% CO 2 for 48 h. ( A–G ) The cells were fixed and stained with Alexa Fluor 488 phalloidin (green) and DAPI (blue). The cells were imaged on a Zeiss <t>LSM</t> 710 confocal microscope using a 40× oil objective and Alexa Fluor 488 and DAPI-specific lasers (488 nm multiline argon and 405 nm diode).
    Scanning Electron Microscope, supplied by Hitachi Ltd, used in various techniques. Bioz Stars score: 95/100, based on 8594 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    JEOL scanning electron microscopy
    Optochemical activation of Eg5 siRNA in HeLa cells. HeLa cells were transfected with caged and non-caged siRNAs (40 pmol). The cells were irradiated (5 min, 25 W, 365 nm) and incubated at 37°C, 5% CO 2 for 48 h. ( A–G ) The cells were fixed and stained with Alexa Fluor 488 phalloidin (green) and DAPI (blue). The cells were imaged on a Zeiss <t>LSM</t> 710 confocal microscope using a 40× oil objective and Alexa Fluor 488 and DAPI-specific lasers (488 nm multiline argon and 405 nm diode).
    Scanning Electron Microscopy, supplied by JEOL, used in various techniques. Bioz Stars score: 95/100, based on 10131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Hitachi Ltd scanning electron microscopy
    <t>SEM</t> images and EDS analysis of sintered ceramics. SEM images of sintered ceramics: ( a ) <t>AZO:Y</t> 0 , ( b ) AZO:Y 0.1 , ( c ) AZO:Y 0.15 , and ( d ) AZO:Y 0.2 . EDS analysis of a ( e ) white ZnAl 2 O 4 particle and ( f ) AZO:Y grain in the AZO:Y 0.2 ceramic sample.
    Scanning Electron Microscopy, supplied by Hitachi Ltd, used in various techniques. Bioz Stars score: 95/100, based on 9799 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Carl Zeiss fluorescence microscopy
    <t>SEM</t> images and EDS analysis of sintered ceramics. SEM images of sintered ceramics: ( a ) <t>AZO:Y</t> 0 , ( b ) AZO:Y 0.1 , ( c ) AZO:Y 0.15 , and ( d ) AZO:Y 0.2 . EDS analysis of a ( e ) white ZnAl 2 O 4 particle and ( f ) AZO:Y grain in the AZO:Y 0.2 ceramic sample.
    Fluorescence Microscopy, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 92/100, based on 14871 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Olympus fv1000 confocal microscope
    IIS and HSF-1 pathways regulated mutant ataxin-3-mediated proteotoxicity in C. elegans neurons. ( A ) Flattened z-series of AT3q130 and daf-2 ; AT3q130 animals that were grown at 15°C for 4 days and transferred to 20°C for 24 h. ( B – E ) Animals were grown at 20°C during their lifespan. White squares highlight the area where the decrease in aggregation is most clear. (A, B and F ) daf-2(e1370) and age-1(hx546) mutations tend to reduce AT3q130-mediated aggregation. The absence of DAF-16 (D) and HSF-1 (E) increased aggregation of mutant ATXN3 (F). The daf-16(mu86) mutation caused a mild aggravation of the aggregation phenotype, visible at day 3 (post-hatching) (D, square). hsf-1(sy441) (E) mutation had a great impact on aggregation, with some aggregates visible already in embryos (arrow). Scale bar, 50 µm. All pictures were obtained using an Olympus <t>FV1000</t> confocal microscope. (F) Quantification of the number of aggregates per area of animal of all strains. Data show the mean ± SD of eight or more animals. Asterisk indicates the significant mean difference between either hsf-1 ; AT3q130 or daf-16 ; AT3q130 and AT3q130 animals; hash symbol indicates the significant difference between hsf-1 ; AT3q130 and daf-16 ; AT3q130 ( ANOVA , applying Bonferroni correction with 95% confidence intervals; * ,# P
    Fv1000 Confocal Microscope, supplied by Olympus, used in various techniques. Bioz Stars score: 95/100, based on 12152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of evolved mutations in the pbpA operon on cell morphology. Cells were grown in DM250 with or without amdinocillin (Mecillinam) and observed using a Zeiss Axioplan 2 microscope. All photographs were obtained using a magnification of ×1,000,

    Journal:

    Article Title: Evolution of Penicillin-Binding Protein 2 Concentration and Cell Shape during a Long-Term Experiment with Escherichia coli ▿

    doi: 10.1128/JB.01419-08

    Figure Lengend Snippet: Effects of evolved mutations in the pbpA operon on cell morphology. Cells were grown in DM250 with or without amdinocillin (Mecillinam) and observed using a Zeiss Axioplan 2 microscope. All photographs were obtained using a magnification of ×1,000,

    Article Snippet: Each strain was grown in DM250 to mid-exponential phase (optical density at 600 nm [OD600 ], 0.2), and cells were observed using a Zeiss Axioplan 2 microscope.

    Techniques: Microscopy

    Localization of overexpressed torsinA proteins in hippocampal neurons of rats and ΔE-torsinA knock-in mice. Cultured neurons were transfected with a construct encoding either WT-torsinA-GFP ( A ) or ΔE-torsinA-GFP ( B ). A modification of the calcium-phosphate method in Fig. 1 was used to enhance the efficiency of transfection ( Jiang and Chen, 2006 ). A confocal laser-scanning microscope was used to observe the neurons. Neurons are shown both as single optical sections at the somatic level (Single) and as maximum intensity projection (MIP) images that represent the overall structure at all heights. Neurons were cultured from WT rats (top row), WT mice (2 nd row), heterozygous (HET, Tor1a +/ΔE , 3 rd row) or homozygous (HOM, Tor1a ΔE/ΔE , bottom row) ΔE-torsinA knock-in mice. Arrows point to representative examples of the diffuse distribution of WT-torsinA-GFP in the cytoplasm ( A ), and ΔE-torsinA-GFP in cytoplasmic inclusion bodies ( B ).

    Journal: bioRxiv

    Article Title: Localization of the signal of dystonia-associated protein torsinA near the Golgi apparatus in cultured central neurons

    doi: 10.1101/2019.12.11.872804

    Figure Lengend Snippet: Localization of overexpressed torsinA proteins in hippocampal neurons of rats and ΔE-torsinA knock-in mice. Cultured neurons were transfected with a construct encoding either WT-torsinA-GFP ( A ) or ΔE-torsinA-GFP ( B ). A modification of the calcium-phosphate method in Fig. 1 was used to enhance the efficiency of transfection ( Jiang and Chen, 2006 ). A confocal laser-scanning microscope was used to observe the neurons. Neurons are shown both as single optical sections at the somatic level (Single) and as maximum intensity projection (MIP) images that represent the overall structure at all heights. Neurons were cultured from WT rats (top row), WT mice (2 nd row), heterozygous (HET, Tor1a +/ΔE , 3 rd row) or homozygous (HOM, Tor1a ΔE/ΔE , bottom row) ΔE-torsinA knock-in mice. Arrows point to representative examples of the diffuse distribution of WT-torsinA-GFP in the cytoplasm ( A ), and ΔE-torsinA-GFP in cytoplasmic inclusion bodies ( B ).

    Article Snippet: For quantitative characterization, the cells were imaged using a confocal laser-scanning microscope (510 confocal, Carl Zeiss MicroImaging GmbH, Göttingen, Germany) equipped with an inverted microscope (Axiovert 100M, Carl Zeiss) and a 40x objective lens (Plan-Neofluor, numerical aperture 1.30).

    Techniques: Knock-In, Mouse Assay, Cell Culture, Transfection, Construct, Modification, Laser-Scanning Microscopy

    Confocal laser scanning microscopy images of DiD-loaded vc378 distribution in the microneedle. Representative confocal images of microneedles coated with DiD-loaded vc378 (left, DiD-loaded vc378; middle, DIC; right, merge). The scale bars are 200 μm.

    Journal: European Journal of Pharmaceutics and Biopharmaceutics

    Article Title: Patchless administration of canine influenza vaccine on dog’s ear using insertion-responsive microneedles (IRMN) without removal of hair and its in vivo efficacy evaluation

    doi: 10.1016/j.ejpb.2020.06.006

    Figure Lengend Snippet: Confocal laser scanning microscopy images of DiD-loaded vc378 distribution in the microneedle. Representative confocal images of microneedles coated with DiD-loaded vc378 (left, DiD-loaded vc378; middle, DIC; right, merge). The scale bars are 200 μm.

    Article Snippet: The cells were then washed with PBS and fluorescent images were collected using a laser scanning confocal microscope (LSM700, Carl Zeiss, Jena, Germany).

    Techniques: Confocal Laser Scanning Microscopy

    Anti-Neu humoral response following rV- neu T vaccination. Panel A : p185 Neu expression in NIH3T3 cells encoding normal rat Neu (LTR-Neu) and in Neu-overexpressing BALB- neu T salivary gland cancer (SALTO) cells by western blotting. A specific polyclonal anti-Neu (C18) antibody was used. NIH3T3 cells were used as negative control. Panel B : Igs from rV- neu T vaccinated mice recognize p185 Neu expressed on the surface of SALTO tumor cells. Immunofluorescence was performed using purified Igs (1μg/ml) of BALB- neu T mice vaccinated with rV- neu T or V-wt. The specific monoclonal antibody anti-Neu Ab4 was used as positive control. Olympus BX51 microscope and I.A.S. version 007 000 Deconvolution 2D software were used to deconvolve z series images of stained native cells. Original magnification x400. Panel C : Serum antibody response of mice upon vaccination with rV- neu T. Numbering identifies immune sera of individual mice. Mouse pre-immune or immune sera were collected prior to vaccination or one week after the second immunization and employed in immunoprecipitation of Neu from LTR-Neu or SALTO tumor cells. p185 Neu specificity was visualized by immunoblotting analysis using receptor-specific polyclonal antibody of immunoprecipitates and compared to direct immunoblotting of LTR-Neu lysate as positive control, or NIH3T3 as negative control.

    Journal: Journal of Translational Medicine

    Article Title: Intratumoral delivery of recombinant vaccinia virus encoding for ErbB2/Neu inhibits the growth of salivary gland carcinoma cells

    doi: 10.1186/1479-5876-12-122

    Figure Lengend Snippet: Anti-Neu humoral response following rV- neu T vaccination. Panel A : p185 Neu expression in NIH3T3 cells encoding normal rat Neu (LTR-Neu) and in Neu-overexpressing BALB- neu T salivary gland cancer (SALTO) cells by western blotting. A specific polyclonal anti-Neu (C18) antibody was used. NIH3T3 cells were used as negative control. Panel B : Igs from rV- neu T vaccinated mice recognize p185 Neu expressed on the surface of SALTO tumor cells. Immunofluorescence was performed using purified Igs (1μg/ml) of BALB- neu T mice vaccinated with rV- neu T or V-wt. The specific monoclonal antibody anti-Neu Ab4 was used as positive control. Olympus BX51 microscope and I.A.S. version 007 000 Deconvolution 2D software were used to deconvolve z series images of stained native cells. Original magnification x400. Panel C : Serum antibody response of mice upon vaccination with rV- neu T. Numbering identifies immune sera of individual mice. Mouse pre-immune or immune sera were collected prior to vaccination or one week after the second immunization and employed in immunoprecipitation of Neu from LTR-Neu or SALTO tumor cells. p185 Neu specificity was visualized by immunoblotting analysis using receptor-specific polyclonal antibody of immunoprecipitates and compared to direct immunoblotting of LTR-Neu lysate as positive control, or NIH3T3 as negative control.

    Article Snippet: After washes with cold PBS, cells were labelled with goat anti-mouse IgG Alexa fluor-488-conjugated antibody (Life Technologies™ Molecular Probes, Oregon, USA) for 1 hour on ice, washed and immediately observed with an Olympus BX51 microscope; I.A.S. version 007 000 Deconvolution 2D software was used to deconvolve z series images of stained native cells [ ].

    Techniques: Expressing, Western Blot, Negative Control, Mouse Assay, Immunofluorescence, Purification, Positive Control, Microscopy, Software, Staining, Immunoprecipitation

    The gene expression of HDL-R in the leg muscle measured by in situ hybridization. Five to ten slides for each tissue were prepared, and images were taken using a binocular microscope (Olympus BX5; Olympus, Japan) coupled to a digital camera (Nikon H550L, Japan). The four images shown in Figure 1 were magnified 400 times. The in situ hybridization images show that the 12:1 and 9:1 groups had lower levels of labeling than the 6:1 and 3:1 groups. The positive rates of HDL-R expression in the 12:1 and 9:1 groups were significantly lower (p

    Journal: Asian-Australasian Journal of Animal Sciences

    Article Title: Lower ω-6/ω-3 Polyunsaturated Fatty Acid Ratios Decrease Fat Deposition by Inhibiting Fat Synthesis in Gosling

    doi: 10.5713/ajas.15.1056

    Figure Lengend Snippet: The gene expression of HDL-R in the leg muscle measured by in situ hybridization. Five to ten slides for each tissue were prepared, and images were taken using a binocular microscope (Olympus BX5; Olympus, Japan) coupled to a digital camera (Nikon H550L, Japan). The four images shown in Figure 1 were magnified 400 times. The in situ hybridization images show that the 12:1 and 9:1 groups had lower levels of labeling than the 6:1 and 3:1 groups. The positive rates of HDL-R expression in the 12:1 and 9:1 groups were significantly lower (p

    Article Snippet: Samples were examined with an OLYMPUS microscope at 400×.

    Techniques: Expressing, In Situ Hybridization, Microscopy, Labeling

    The gene expression of LDL-R in the leg muscle measured by in situ hybridization. Five to ten slides for each tissue were prepared, and images were taken using binocular microscope (Olympus BX5; Olympus, Japan) coupled to a digital camera (Nikon H550L, Japan). The four images shown in Figure 2 were all magnified 400. Figure 2 shows that the positive rates of LDL-R expression between the different groups had a tendency to decrease with decreasing ω-6/ω-3 polyunsaturated fatty acid ratios (data shown in Table 2 ).

    Journal: Asian-Australasian Journal of Animal Sciences

    Article Title: Lower ω-6/ω-3 Polyunsaturated Fatty Acid Ratios Decrease Fat Deposition by Inhibiting Fat Synthesis in Gosling

    doi: 10.5713/ajas.15.1056

    Figure Lengend Snippet: The gene expression of LDL-R in the leg muscle measured by in situ hybridization. Five to ten slides for each tissue were prepared, and images were taken using binocular microscope (Olympus BX5; Olympus, Japan) coupled to a digital camera (Nikon H550L, Japan). The four images shown in Figure 2 were all magnified 400. Figure 2 shows that the positive rates of LDL-R expression between the different groups had a tendency to decrease with decreasing ω-6/ω-3 polyunsaturated fatty acid ratios (data shown in Table 2 ).

    Article Snippet: Samples were examined with an OLYMPUS microscope at 400×.

    Techniques: Expressing, In Situ Hybridization, Microscopy

    Heterozygosity for Cldn14 increases tumour blood vessel leakage and decreases intratumoural hypoxia. Wild-type, Cldn14-heterozygous and Cldn14-null mice were injected subcutaneously in the flank with 0.5×10 6 B16F10 melanoma or Lewis Lung Carcinoma (LLC) cells. ( A ) At 10 days post inoculation, PE-conjugated anti-PECAM antibody and Hoechst dye were injected via the tail vein prior to sacrifice. Midline sections (100 µm) of snap-frozen tumours were fixed, mounted and imaged using a Zeiss LSM 510 confocal microscope. The extent of Hoechst leakage was measured in z-stacks using ImageJ. Bars show mean Hoechst leakage relative to PECAM signal ± SEM. Blood vessel leakage is increased significantly in Cldn14-het mice when compared with WT and Cldn14-null mice. ( B ) Representative images of Hoechst (blue) and PECAM (red) detection. ( C ) Tumour-bearing mice from each genotype were injected with pimonidazole prior to sacrifice to measure hypoxic areas within the tumour. 8 µm tumour cryosections were then double stained with anti-pimonidazole antibody (green) to highlight hypoxic areas and anti-PECAM antibody to identify blood vessels. The hypoxic index was quantified relative to PECAM staining using image J software. Bars represent mean relative hypoxic index ± SEM. ( D ) Representative images of pimonidazole detection and PECAM-positive blood vessels in tumour sections. Arrows , blood vessels; Asterisks , pimonidazole-positive staining. Scale bars: A 50 µm; D 200 µm. N = 4 tumours per genotype. NSD: not statistically different, * P

    Journal: PLoS ONE

    Article Title: Stromal Claudin14-Heterozygosity, but Not Deletion, Increases Tumour Blood Leakage without Affecting Tumour Growth

    doi: 10.1371/journal.pone.0062516

    Figure Lengend Snippet: Heterozygosity for Cldn14 increases tumour blood vessel leakage and decreases intratumoural hypoxia. Wild-type, Cldn14-heterozygous and Cldn14-null mice were injected subcutaneously in the flank with 0.5×10 6 B16F10 melanoma or Lewis Lung Carcinoma (LLC) cells. ( A ) At 10 days post inoculation, PE-conjugated anti-PECAM antibody and Hoechst dye were injected via the tail vein prior to sacrifice. Midline sections (100 µm) of snap-frozen tumours were fixed, mounted and imaged using a Zeiss LSM 510 confocal microscope. The extent of Hoechst leakage was measured in z-stacks using ImageJ. Bars show mean Hoechst leakage relative to PECAM signal ± SEM. Blood vessel leakage is increased significantly in Cldn14-het mice when compared with WT and Cldn14-null mice. ( B ) Representative images of Hoechst (blue) and PECAM (red) detection. ( C ) Tumour-bearing mice from each genotype were injected with pimonidazole prior to sacrifice to measure hypoxic areas within the tumour. 8 µm tumour cryosections were then double stained with anti-pimonidazole antibody (green) to highlight hypoxic areas and anti-PECAM antibody to identify blood vessels. The hypoxic index was quantified relative to PECAM staining using image J software. Bars represent mean relative hypoxic index ± SEM. ( D ) Representative images of pimonidazole detection and PECAM-positive blood vessels in tumour sections. Arrows , blood vessels; Asterisks , pimonidazole-positive staining. Scale bars: A 50 µm; D 200 µm. N = 4 tumours per genotype. NSD: not statistically different, * P

    Article Snippet: 100 µm Z-stacks (stack interval 0.5 µm, 20× magnification) were taken using a Zeiss LSM 510 confocal microscope.

    Techniques: Mouse Assay, Injection, Microscopy, Staining, Software

    CpG-A and CpG-B are distributed in different compartments in PDCs. Purified PDCs were cultured with fluorescent CpG-A (A and C) or CpG-B (E and G). Cells were fixed, stained intracellularly with (A and E) antitransferrin receptor (TfR) or (C and G) anti–LAMP-1 antibodies, and imaged by confocal microscopy. Images were acquired using a ZEISS LSM 510 META confocal microscope. We used CpG-A-Rhodamine green-X and CpG-B-Alexa488. Intensity profiles of the merged channel along three randomly chosen lines ( 1 , 2 , and 3 shown on each merged staining) were analyzed using the profile tools of the Zeiss LSM software. Examples are shown for (B) CpG-A and TfR, (D) CpG-A and LAMP-1, (F) CpG-B and TfR, and (H) CpG-B and LAMP1. The green line represents the intensity of the ISS, whereas the red line represents the intensity of the endosomal marker. Overlap of the two profiles indicates spatial correlation for the occurrence of the two fluorescent signals. Representative data of 5–10 individual donors are shown.

    Journal: The Journal of Experimental Medicine

    Article Title: Properties regulating the nature of the plasmacytoid dendritic cell response to Toll-like receptor 9 activation

    doi: 10.1084/jem.20060401

    Figure Lengend Snippet: CpG-A and CpG-B are distributed in different compartments in PDCs. Purified PDCs were cultured with fluorescent CpG-A (A and C) or CpG-B (E and G). Cells were fixed, stained intracellularly with (A and E) antitransferrin receptor (TfR) or (C and G) anti–LAMP-1 antibodies, and imaged by confocal microscopy. Images were acquired using a ZEISS LSM 510 META confocal microscope. We used CpG-A-Rhodamine green-X and CpG-B-Alexa488. Intensity profiles of the merged channel along three randomly chosen lines ( 1 , 2 , and 3 shown on each merged staining) were analyzed using the profile tools of the Zeiss LSM software. Examples are shown for (B) CpG-A and TfR, (D) CpG-A and LAMP-1, (F) CpG-B and TfR, and (H) CpG-B and LAMP1. The green line represents the intensity of the ISS, whereas the red line represents the intensity of the endosomal marker. Overlap of the two profiles indicates spatial correlation for the occurrence of the two fluorescent signals. Representative data of 5–10 individual donors are shown.

    Article Snippet: Images were acquired using a ZEISS LSM 510 META confocal microscope and a 63×/1.4 N.A. objective, with the pinhole set for a section thickness of 0.8 μm (pinhole set to 1 airy unit in each channels).

    Techniques: Purification, Cell Culture, Staining, Confocal Microscopy, Microscopy, Software, Marker

    The secondary structure of ISS is regulating their intracellular localization in human PDCs. (A and B) Purified PDCs were cultured with fluorescent ISS for 90 min. Cells were fixed, stained intracellularly with antitransferrin receptor (TfR) or anti-LAMP1 antibodies, and imaged by confocal microscopy. We used CpG-A ss-Rhodamine green-X from the same preparation as CpG-A used in Fig. 4 and CpG-B-Alexa488 premixed with PMXB for 30 min. Images were acquired using a ZEISS LSM 510 META confocal microscope. (B) Between 100 and 200 cells were analyzed from three donors for colocalization between the ODN and either transferrin receptor (TfR) or LAMP-1 (LP1).

    Journal: The Journal of Experimental Medicine

    Article Title: Properties regulating the nature of the plasmacytoid dendritic cell response to Toll-like receptor 9 activation

    doi: 10.1084/jem.20060401

    Figure Lengend Snippet: The secondary structure of ISS is regulating their intracellular localization in human PDCs. (A and B) Purified PDCs were cultured with fluorescent ISS for 90 min. Cells were fixed, stained intracellularly with antitransferrin receptor (TfR) or anti-LAMP1 antibodies, and imaged by confocal microscopy. We used CpG-A ss-Rhodamine green-X from the same preparation as CpG-A used in Fig. 4 and CpG-B-Alexa488 premixed with PMXB for 30 min. Images were acquired using a ZEISS LSM 510 META confocal microscope. (B) Between 100 and 200 cells were analyzed from three donors for colocalization between the ODN and either transferrin receptor (TfR) or LAMP-1 (LP1).

    Article Snippet: Images were acquired using a ZEISS LSM 510 META confocal microscope and a 63×/1.4 N.A. objective, with the pinhole set for a section thickness of 0.8 μm (pinhole set to 1 airy unit in each channels).

    Techniques: Purification, Cell Culture, Staining, Confocal Microscopy, Microscopy

    Stokes-shift and anti-Stokes-shift luminescence images with scale bars of 20 μm at 40× magnification of Hela cells together with 10 μM of probes A and B in pH buffers from 2.5, 3.5, 4.5, 5.5, 6.5 to 7.5 holding 5 mg/L nigericin. Stokes-shift luminescence image (red-color) was acquired under 633 nm excitation with HeNe continuous wave laser while anti-Stokes-shift luminescence image (green-color) was obtained under 800 nm excitationt with a pulse laser. Both Stokes-shift and anti-Stokes-shift luminescence were collected from 650 to 700 nm by using Zeiss LSM510 confocal fluorescence microscope.

    Journal: ACS applied bio materials

    Article Title: New Near-infrared Fluorescent Probes with Single-photon Anti-Stokes-shift Fluorescence for Sensitive Determination of pH Variances in Lysosomes with a Double-Checked Capability

    doi: 10.1021/acsabm.8b00020

    Figure Lengend Snippet: Stokes-shift and anti-Stokes-shift luminescence images with scale bars of 20 μm at 40× magnification of Hela cells together with 10 μM of probes A and B in pH buffers from 2.5, 3.5, 4.5, 5.5, 6.5 to 7.5 holding 5 mg/L nigericin. Stokes-shift luminescence image (red-color) was acquired under 633 nm excitation with HeNe continuous wave laser while anti-Stokes-shift luminescence image (green-color) was obtained under 800 nm excitationt with a pulse laser. Both Stokes-shift and anti-Stokes-shift luminescence were collected from 650 to 700 nm by using Zeiss LSM510 confocal fluorescence microscope.

    Article Snippet: We further used the probes to investigate pH changes in lysosomes with high-resolution using a Zeiss LSM510 confocal microscope (Figures , , ).

    Techniques: Fluorescence, Microscopy

    Stokes-shift and anti-Stokes-shift luminescence images with scale bars of 20 μm at 40× magnification of KB cells together with 10 μM of probes A and B in pH buffers from 2.5, 3.5, 4.5, 5.5, 6.5 to 7.5 holding 5 mg/L nigericin. Stokes-shift luminescence image (red-color) was acquired under 633 nm excitation with a HeNe continuous wave laser while anti-Stokes-shift luminescence image (green-color) was obtained under 800 nm excitation with a pulse laser. Both Stokes-shift and anti-Stokes-shift luminescence were collected from 650 to 700 nm by Zeiss LSM510 confocal fluorescence microscope.

    Journal: ACS applied bio materials

    Article Title: New Near-infrared Fluorescent Probes with Single-photon Anti-Stokes-shift Fluorescence for Sensitive Determination of pH Variances in Lysosomes with a Double-Checked Capability

    doi: 10.1021/acsabm.8b00020

    Figure Lengend Snippet: Stokes-shift and anti-Stokes-shift luminescence images with scale bars of 20 μm at 40× magnification of KB cells together with 10 μM of probes A and B in pH buffers from 2.5, 3.5, 4.5, 5.5, 6.5 to 7.5 holding 5 mg/L nigericin. Stokes-shift luminescence image (red-color) was acquired under 633 nm excitation with a HeNe continuous wave laser while anti-Stokes-shift luminescence image (green-color) was obtained under 800 nm excitation with a pulse laser. Both Stokes-shift and anti-Stokes-shift luminescence were collected from 650 to 700 nm by Zeiss LSM510 confocal fluorescence microscope.

    Article Snippet: We further used the probes to investigate pH changes in lysosomes with high-resolution using a Zeiss LSM510 confocal microscope (Figures , , ).

    Techniques: Fluorescence, Microscopy

    PDGF-AA-induced dorsal ruffle formation is diminished in WIP −/− fibroblasts. a Control (WIP +/+ ) and WIP −/− primary murine fibroblasts were serum starved over night (0 min) or serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and FITC-secondary antibody and imaged in a Zeiss microscope to identify dorsal ruffles (white arrow). Magnifications of the boxed areas are shown as right panels. b The percentage of WIP +/+ (black) and WIP −/− (white) cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. c WIP −/− primary fibroblasts were lentivirally transduced to express control cherry or WIP-cherry, starved and incubated with PDGF-AA for 8 or 15 min. Fixed cells were imaged. d The percentage of WIP −/− cells expressing cherry (white) or WIP-cherry (black) and forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. e Representative western blot of PDGFRα expression in soluble lysates of lung-derived fibroblasts from WIP +/+ and WIP −/− mice. Numbers indicate relative expression levels of the protein to control fibroblasts determined by densitometry. ß-actin labeling confirmed equivalent protein loading control. f WIP +/+ (black) and WIP −/− (white) primary fibroblasts were grown in the presence of serum (left panel) and stained with anti-PDGFRα plus labeled secondary antibody and analysed by FACS. The mean fluorescence intensity of positive cells in both populations is represented. Right panel, WIP +/+ (black) and WIP −/− (white) primary fibroblasts were starved (time 0) and stimulated with PDGF-AA and stained as above. Percent of mean fluorescence intensity relative to starvation is represented. Arbitrary units (a.u.). * p

    Journal: PLoS ONE

    Article Title: WIP Regulates Persistence of Cell Migration and Ruffle Formation in Both Mesenchymal and Amoeboid Modes of Motility

    doi: 10.1371/journal.pone.0070364

    Figure Lengend Snippet: PDGF-AA-induced dorsal ruffle formation is diminished in WIP −/− fibroblasts. a Control (WIP +/+ ) and WIP −/− primary murine fibroblasts were serum starved over night (0 min) or serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and FITC-secondary antibody and imaged in a Zeiss microscope to identify dorsal ruffles (white arrow). Magnifications of the boxed areas are shown as right panels. b The percentage of WIP +/+ (black) and WIP −/− (white) cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. c WIP −/− primary fibroblasts were lentivirally transduced to express control cherry or WIP-cherry, starved and incubated with PDGF-AA for 8 or 15 min. Fixed cells were imaged. d The percentage of WIP −/− cells expressing cherry (white) or WIP-cherry (black) and forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. e Representative western blot of PDGFRα expression in soluble lysates of lung-derived fibroblasts from WIP +/+ and WIP −/− mice. Numbers indicate relative expression levels of the protein to control fibroblasts determined by densitometry. ß-actin labeling confirmed equivalent protein loading control. f WIP +/+ (black) and WIP −/− (white) primary fibroblasts were grown in the presence of serum (left panel) and stained with anti-PDGFRα plus labeled secondary antibody and analysed by FACS. The mean fluorescence intensity of positive cells in both populations is represented. Right panel, WIP +/+ (black) and WIP −/− (white) primary fibroblasts were starved (time 0) and stimulated with PDGF-AA and stained as above. Percent of mean fluorescence intensity relative to starvation is represented. Arbitrary units (a.u.). * p

    Article Snippet: Fixed and permeabilised cells were stained with FITC-phalloidin to label actin filaments and imaged in a Zeiss microscope. (PPTX) Click here for additional data file.

    Techniques: Staining, Microscopy, Incubation, Expressing, Western Blot, Derivative Assay, Mouse Assay, Labeling, FACS, Fluorescence

    Nck and N-WASP binding to WIP contribute to dorsal ruffle formation induced by PDGF-AA stimulation. a WIP −/− primary murine fibroblasts were transduced with recombinant lentivirus coding for GFP, WIP-GFP, WIPΔNBD-GFP (missing the Nck binding domain) or WIPΔWBD-GFP (missing the N-WASP binding site). Soluble protein extracts were subjected to western blot analysis, using anti-GFP as probe, and developed with ECL detection kit. b Transduced WIP −/− murine fibroblasts were serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and FITC-secondary antibody and imaged in a Zeiss microscope. The percentage of GFP-positive cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. One way ANOVA test and Test of Tukey * p

    Journal: PLoS ONE

    Article Title: WIP Regulates Persistence of Cell Migration and Ruffle Formation in Both Mesenchymal and Amoeboid Modes of Motility

    doi: 10.1371/journal.pone.0070364

    Figure Lengend Snippet: Nck and N-WASP binding to WIP contribute to dorsal ruffle formation induced by PDGF-AA stimulation. a WIP −/− primary murine fibroblasts were transduced with recombinant lentivirus coding for GFP, WIP-GFP, WIPΔNBD-GFP (missing the Nck binding domain) or WIPΔWBD-GFP (missing the N-WASP binding site). Soluble protein extracts were subjected to western blot analysis, using anti-GFP as probe, and developed with ECL detection kit. b Transduced WIP −/− murine fibroblasts were serum starved and stimulated with PDGF-AA for increasing times (8 and 15 min). Fixed and permeabilised cells were stained with anti-cortactin and FITC-secondary antibody and imaged in a Zeiss microscope. The percentage of GFP-positive cells forming dorsal ruffles after PDGF-AA stimulation is plotted against incubation times. One way ANOVA test and Test of Tukey * p

    Article Snippet: Fixed and permeabilised cells were stained with FITC-phalloidin to label actin filaments and imaged in a Zeiss microscope. (PPTX) Click here for additional data file.

    Techniques: Binding Assay, Transduction, Recombinant, Western Blot, Staining, Microscopy, Incubation

    Localization of Mel-AF on A375 cells was analyzed using a confocal laser scanning microscope. The cells were treated with FITC-conjugated Mel-AF for 2 h. ( A ) Confocal images show the localization of Mel-AF traced with FITC (green). The membrane compartment was indicated by staining with a specific antibody to CD46, a membrane cofactor protein followed by Alexa Fluor 568 rabbit anti-mouse IgG (red). The cellular nuclei were counterstained by DAPI (blue). ( B ) Orthogonal imaging analysis was performed to confirm the localization of Mel-AF on the cell membrane (yellow). Scale bar, 10 μm.

    Journal: Antibiotics

    Article Title: Melittin from Apis florea Venom as a Promising Therapeutic Agent for Skin Cancer Treatment

    doi: 10.3390/antibiotics9080517

    Figure Lengend Snippet: Localization of Mel-AF on A375 cells was analyzed using a confocal laser scanning microscope. The cells were treated with FITC-conjugated Mel-AF for 2 h. ( A ) Confocal images show the localization of Mel-AF traced with FITC (green). The membrane compartment was indicated by staining with a specific antibody to CD46, a membrane cofactor protein followed by Alexa Fluor 568 rabbit anti-mouse IgG (red). The cellular nuclei were counterstained by DAPI (blue). ( B ) Orthogonal imaging analysis was performed to confirm the localization of Mel-AF on the cell membrane (yellow). Scale bar, 10 μm.

    Article Snippet: Images were acquired using a confocal laser scanning microscope (FluoView FV1000, Olympus).

    Techniques: Laser-Scanning Microscopy, Staining, Imaging

    Optochemical activation of Eg5 siRNA in HeLa cells. HeLa cells were transfected with caged and non-caged siRNAs (40 pmol). The cells were irradiated (5 min, 25 W, 365 nm) and incubated at 37°C, 5% CO 2 for 48 h. ( A–G ) The cells were fixed and stained with Alexa Fluor 488 phalloidin (green) and DAPI (blue). The cells were imaged on a Zeiss LSM 710 confocal microscope using a 40× oil objective and Alexa Fluor 488 and DAPI-specific lasers (488 nm multiline argon and 405 nm diode).

    Journal: Nucleic Acids Research

    Article Title: Optochemical control of RNA interference in mammalian cells

    doi: 10.1093/nar/gkt806

    Figure Lengend Snippet: Optochemical activation of Eg5 siRNA in HeLa cells. HeLa cells were transfected with caged and non-caged siRNAs (40 pmol). The cells were irradiated (5 min, 25 W, 365 nm) and incubated at 37°C, 5% CO 2 for 48 h. ( A–G ) The cells were fixed and stained with Alexa Fluor 488 phalloidin (green) and DAPI (blue). The cells were imaged on a Zeiss LSM 710 confocal microscope using a 40× oil objective and Alexa Fluor 488 and DAPI-specific lasers (488 nm multiline argon and 405 nm diode).

    Article Snippet: Cells were imaged on a Zeiss LSM 710 confocal microscope (40× oil objective).

    Techniques: Activation Assay, Transfection, Irradiation, Incubation, Staining, Microscopy

    SEM images and EDS analysis of sintered ceramics. SEM images of sintered ceramics: ( a ) AZO:Y 0 , ( b ) AZO:Y 0.1 , ( c ) AZO:Y 0.15 , and ( d ) AZO:Y 0.2 . EDS analysis of a ( e ) white ZnAl 2 O 4 particle and ( f ) AZO:Y grain in the AZO:Y 0.2 ceramic sample.

    Journal: Nanoscale Research Letters

    Article Title: Nearly full-dense and fine-grained AZO:Y ceramics sintered from the corresponding nanoparticles

    doi: 10.1186/1556-276X-7-481

    Figure Lengend Snippet: SEM images and EDS analysis of sintered ceramics. SEM images of sintered ceramics: ( a ) AZO:Y 0 , ( b ) AZO:Y 0.1 , ( c ) AZO:Y 0.15 , and ( d ) AZO:Y 0.2 . EDS analysis of a ( e ) white ZnAl 2 O 4 particle and ( f ) AZO:Y grain in the AZO:Y 0.2 ceramic sample.

    Article Snippet: The morphology, microstructure, and composition analyses of the AZO:Y nanoparticles and the sintered bodies were performed using a scanning electron microscopy(SEM)/energy-dispersive X-ray analysis (EDAX) system (S-4800, Hitachi Ltd., Tokyo, Japan).

    Techniques:

    SEM images and the calculated particle sizes. Images of AZOY nanoparticles calcined at 600 °C for 2 h: ( a ) AZO:Y 0 , ( b ) AZO:Y 0.1 , ( c ) AZO:Y 0.15 , and ( d ) AZO:Y 0.2 . ( e ) The plot of the particle sizes calculated from ( a to d ) SEM images as a function of Y 2 O 3 content.

    Journal: Nanoscale Research Letters

    Article Title: Nearly full-dense and fine-grained AZO:Y ceramics sintered from the corresponding nanoparticles

    doi: 10.1186/1556-276X-7-481

    Figure Lengend Snippet: SEM images and the calculated particle sizes. Images of AZOY nanoparticles calcined at 600 °C for 2 h: ( a ) AZO:Y 0 , ( b ) AZO:Y 0.1 , ( c ) AZO:Y 0.15 , and ( d ) AZO:Y 0.2 . ( e ) The plot of the particle sizes calculated from ( a to d ) SEM images as a function of Y 2 O 3 content.

    Article Snippet: The morphology, microstructure, and composition analyses of the AZO:Y nanoparticles and the sintered bodies were performed using a scanning electron microscopy(SEM)/energy-dispersive X-ray analysis (EDAX) system (S-4800, Hitachi Ltd., Tokyo, Japan).

    Techniques:

    IIS and HSF-1 pathways regulated mutant ataxin-3-mediated proteotoxicity in C. elegans neurons. ( A ) Flattened z-series of AT3q130 and daf-2 ; AT3q130 animals that were grown at 15°C for 4 days and transferred to 20°C for 24 h. ( B – E ) Animals were grown at 20°C during their lifespan. White squares highlight the area where the decrease in aggregation is most clear. (A, B and F ) daf-2(e1370) and age-1(hx546) mutations tend to reduce AT3q130-mediated aggregation. The absence of DAF-16 (D) and HSF-1 (E) increased aggregation of mutant ATXN3 (F). The daf-16(mu86) mutation caused a mild aggravation of the aggregation phenotype, visible at day 3 (post-hatching) (D, square). hsf-1(sy441) (E) mutation had a great impact on aggregation, with some aggregates visible already in embryos (arrow). Scale bar, 50 µm. All pictures were obtained using an Olympus FV1000 confocal microscope. (F) Quantification of the number of aggregates per area of animal of all strains. Data show the mean ± SD of eight or more animals. Asterisk indicates the significant mean difference between either hsf-1 ; AT3q130 or daf-16 ; AT3q130 and AT3q130 animals; hash symbol indicates the significant difference between hsf-1 ; AT3q130 and daf-16 ; AT3q130 ( ANOVA , applying Bonferroni correction with 95% confidence intervals; * ,# P

    Journal: Human Molecular Genetics

    Article Title: Neuron-specific proteotoxicity of mutant ataxin-3 in C. elegans: rescue by the DAF-16 and HSF-1 pathways

    doi: 10.1093/hmg/ddr203

    Figure Lengend Snippet: IIS and HSF-1 pathways regulated mutant ataxin-3-mediated proteotoxicity in C. elegans neurons. ( A ) Flattened z-series of AT3q130 and daf-2 ; AT3q130 animals that were grown at 15°C for 4 days and transferred to 20°C for 24 h. ( B – E ) Animals were grown at 20°C during their lifespan. White squares highlight the area where the decrease in aggregation is most clear. (A, B and F ) daf-2(e1370) and age-1(hx546) mutations tend to reduce AT3q130-mediated aggregation. The absence of DAF-16 (D) and HSF-1 (E) increased aggregation of mutant ATXN3 (F). The daf-16(mu86) mutation caused a mild aggravation of the aggregation phenotype, visible at day 3 (post-hatching) (D, square). hsf-1(sy441) (E) mutation had a great impact on aggregation, with some aggregates visible already in embryos (arrow). Scale bar, 50 µm. All pictures were obtained using an Olympus FV1000 confocal microscope. (F) Quantification of the number of aggregates per area of animal of all strains. Data show the mean ± SD of eight or more animals. Asterisk indicates the significant mean difference between either hsf-1 ; AT3q130 or daf-16 ; AT3q130 and AT3q130 animals; hash symbol indicates the significant difference between hsf-1 ; AT3q130 and daf-16 ; AT3q130 ( ANOVA , applying Bonferroni correction with 95% confidence intervals; * ,# P

    Article Snippet: Caenorhabditis elegans fluorescent images were acquired using the Olympus FV1000 confocal microscope.

    Techniques: Mutagenesis, Microscopy