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  • 98
    Carl Zeiss fluorescence microscope
    (a) Immunofluorescence staining of the autophagosomal protein LC3 (green fluorescence) and of the lysosomal protein LAMP1 (red fluorescence) and (b) of BECLIN 1 (green fluorescence) and Golgi-associated Golgin97 (red fluorescence) in SKOV3 cells subjected to arginine starvation. Nuclei were labelled with DAPI. Images were captured with <t>ZEISS</t> fluorescence microscope <t>(Axio</t> Imager A1) equipped with Axio Vision Software v. 4.6.3. Magnification 1000x.
    Fluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 98/100, based on 46161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Olympus fluorescence microscope
    MPT0B098 inhibits hypoxia-induced F-actin rearrangement and FAK phosphorylation. a Immunofluorescent analysis of F-actin. OEC-M1 cells were treated with various concentrations, indicated as fold of IC 50 , of MPT0B098 for 18 h under hypoxic conditions, stained with phalloidin to label F-actin (red), counterstained with DAPI (blue), and then observed using an <t>OLYMPUS</t> florescence microscope. b Effect of MPT0B098 on FAK phosphorylation and expression. OEC-M1 cells were treated with various concentrations of MPT0B098 for 18 h.At the end of the drug treatment, cell lysates were prepared and analyzed by SDS-PAGE and Western blot. β-Actin was used as an internal control. c Each bar depicts the mean of the relative intensities of Twist and SNAI2/Slug from three independent experiments (* p
    Fluorescence Microscope, supplied by Olympus, used in various techniques. Bioz Stars score: 99/100, based on 85250 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Carl Zeiss fluorescence microscopy
    MPT0B098 inhibits hypoxia-induced F-actin rearrangement and FAK phosphorylation. a Immunofluorescent analysis of F-actin. OEC-M1 cells were treated with various concentrations, indicated as fold of IC 50 , of MPT0B098 for 18 h under hypoxic conditions, stained with phalloidin to label F-actin (red), counterstained with DAPI (blue), and then observed using an <t>OLYMPUS</t> florescence microscope. b Effect of MPT0B098 on FAK phosphorylation and expression. OEC-M1 cells were treated with various concentrations of MPT0B098 for 18 h.At the end of the drug treatment, cell lysates were prepared and analyzed by SDS-PAGE and Western blot. β-Actin was used as an internal control. c Each bar depicts the mean of the relative intensities of Twist and SNAI2/Slug from three independent experiments (* p
    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 fluorescence microscopy
    MPT0B098 inhibits hypoxia-induced F-actin rearrangement and FAK phosphorylation. a Immunofluorescent analysis of F-actin. OEC-M1 cells were treated with various concentrations, indicated as fold of IC 50 , of MPT0B098 for 18 h under hypoxic conditions, stained with phalloidin to label F-actin (red), counterstained with DAPI (blue), and then observed using an <t>OLYMPUS</t> florescence microscope. b Effect of MPT0B098 on FAK phosphorylation and expression. OEC-M1 cells were treated with various concentrations of MPT0B098 for 18 h.At the end of the drug treatment, cell lysates were prepared and analyzed by SDS-PAGE and Western blot. β-Actin was used as an internal control. c Each bar depicts the mean of the relative intensities of Twist and SNAI2/Slug from three independent experiments (* p
    Fluorescence Microscopy, supplied by Olympus, used in various techniques. Bioz Stars score: 95/100, based on 21746 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher fluorescence microscope
    MPT0B098 inhibits hypoxia-induced F-actin rearrangement and FAK phosphorylation. a Immunofluorescent analysis of F-actin. OEC-M1 cells were treated with various concentrations, indicated as fold of IC 50 , of MPT0B098 for 18 h under hypoxic conditions, stained with phalloidin to label F-actin (red), counterstained with DAPI (blue), and then observed using an <t>OLYMPUS</t> florescence microscope. b Effect of MPT0B098 on FAK phosphorylation and expression. OEC-M1 cells were treated with various concentrations of MPT0B098 for 18 h.At the end of the drug treatment, cell lysates were prepared and analyzed by SDS-PAGE and Western blot. β-Actin was used as an internal control. c Each bar depicts the mean of the relative intensities of Twist and SNAI2/Slug from three independent experiments (* p
    Fluorescence Microscope, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 3208 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Carl Zeiss zeiss fluorescent microscope
    The impact of CuO NMs and CuSO 4 on Caco-2 cell morphology. a ) Following differentiation, cell monolayers were exposed to cell culture medium (control) or 6.34 μg/cm 2 Cu of CuO NMs and CuSO 4 for 24 h and then fixed and stained for the tight junction protein ZO-1 (green) and nucleus with DAPI (blue). The images were obtained with <t>Zeiss</t> <t>LSM880</t> confocal microscope using the Zen program for data analyses. Scale bar = 20 μm. b ) Assessment of cell morphology using SEM. Differentiated and undifferentiated Caco-2 cells were exposed to 12.68 μg/cm 2 Cu of CuO NMs for 24 h and then were washed, fixed, dehydrated, dried and examined by FIB/SEM. i and ii are control differentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. iii and iv are control undifferentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. v and vi are differentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively and vii and viii are undifferentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively
    Zeiss Fluorescent Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 2365 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Carl Zeiss confocal fluorescence microscope
    The impact of CuO NMs and CuSO 4 on Caco-2 cell morphology. a ) Following differentiation, cell monolayers were exposed to cell culture medium (control) or 6.34 μg/cm 2 Cu of CuO NMs and CuSO 4 for 24 h and then fixed and stained for the tight junction protein ZO-1 (green) and nucleus with DAPI (blue). The images were obtained with <t>Zeiss</t> <t>LSM880</t> confocal microscope using the Zen program for data analyses. Scale bar = 20 μm. b ) Assessment of cell morphology using SEM. Differentiated and undifferentiated Caco-2 cells were exposed to 12.68 μg/cm 2 Cu of CuO NMs for 24 h and then were washed, fixed, dehydrated, dried and examined by FIB/SEM. i and ii are control differentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. iii and iv are control undifferentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. v and vi are differentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively and vii and viii are undifferentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively
    Confocal Fluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 2219 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 2219 article reviews
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    94
    Olympus confocal fluorescence microscope
    The impact of CuO NMs and CuSO 4 on Caco-2 cell morphology. a ) Following differentiation, cell monolayers were exposed to cell culture medium (control) or 6.34 μg/cm 2 Cu of CuO NMs and CuSO 4 for 24 h and then fixed and stained for the tight junction protein ZO-1 (green) and nucleus with DAPI (blue). The images were obtained with <t>Zeiss</t> <t>LSM880</t> confocal microscope using the Zen program for data analyses. Scale bar = 20 μm. b ) Assessment of cell morphology using SEM. Differentiated and undifferentiated Caco-2 cells were exposed to 12.68 μg/cm 2 Cu of CuO NMs for 24 h and then were washed, fixed, dehydrated, dried and examined by FIB/SEM. i and ii are control differentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. iii and iv are control undifferentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. v and vi are differentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively and vii and viii are undifferentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively
    Confocal Fluorescence Microscope, supplied by Olympus, used in various techniques. Bioz Stars score: 94/100, based on 2284 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Carl Zeiss axiovert 200 fluorescence microscope
    Indirect immunofluorescence analysis of surface ABCG2, SSEA-1, and SSEA-4 coexpression during differentiation. (A): WA01 cells (control); (B and C): WA01 cells treated with 50 ng/mL or 100 ng/mL of recombinant BMP-4 for 6 days. C1 indicates BMP-4 concentration 1 (i.e., 50 ng/mL) and C2 denotes BMP-4 concentration 2 (i.e., 100 ng/mL). The images were acquired with an <t>Axiovert</t> 200 fluorescence microscope (Zeiss). Bar, 50 μm. (D and E): Quantitative analysis of the coexpression of surface ABCG2 and SSEA-1 in WA01 cells (D) and WA09 cells (E) under the treatment of the cells with 100 ng/mL of BMP-4 for 0 (control), 24, 96, and 144 hours. Mean plasma membrane fluorescence intensity (FI or IF) from a representative pixel per cell was measured by the ImageJ program. The average of the mean fluorescence intensity (FI) and its standard deviation (SD) from 50 individual cells are shown. The Pearson correlations between ABCG2 and SSEA-1 are shown in the right panels. Abbreviations: CE, coefficient; hr, hour(s).
    Axiovert 200 Fluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 2509 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Carl Zeiss confocal fluorescence microscopy
    Indirect immunofluorescence analysis of surface ABCG2, SSEA-1, and SSEA-4 coexpression during differentiation. (A): WA01 cells (control); (B and C): WA01 cells treated with 50 ng/mL or 100 ng/mL of recombinant BMP-4 for 6 days. C1 indicates BMP-4 concentration 1 (i.e., 50 ng/mL) and C2 denotes BMP-4 concentration 2 (i.e., 100 ng/mL). The images were acquired with an <t>Axiovert</t> 200 fluorescence microscope (Zeiss). Bar, 50 μm. (D and E): Quantitative analysis of the coexpression of surface ABCG2 and SSEA-1 in WA01 cells (D) and WA09 cells (E) under the treatment of the cells with 100 ng/mL of BMP-4 for 0 (control), 24, 96, and 144 hours. Mean plasma membrane fluorescence intensity (FI or IF) from a representative pixel per cell was measured by the ImageJ program. The average of the mean fluorescence intensity (FI) and its standard deviation (SD) from 50 individual cells are shown. The Pearson correlations between ABCG2 and SSEA-1 are shown in the right panels. Abbreviations: CE, coefficient; hr, hour(s).
    Confocal Fluorescence Microscopy, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 94/100, based on 2528 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Nikon fluorescence microscope
    Indirect immunofluorescence analysis of surface ABCG2, SSEA-1, and SSEA-4 coexpression during differentiation. (A): WA01 cells (control); (B and C): WA01 cells treated with 50 ng/mL or 100 ng/mL of recombinant BMP-4 for 6 days. C1 indicates BMP-4 concentration 1 (i.e., 50 ng/mL) and C2 denotes BMP-4 concentration 2 (i.e., 100 ng/mL). The images were acquired with an <t>Axiovert</t> 200 fluorescence microscope (Zeiss). Bar, 50 μm. (D and E): Quantitative analysis of the coexpression of surface ABCG2 and SSEA-1 in WA01 cells (D) and WA09 cells (E) under the treatment of the cells with 100 ng/mL of BMP-4 for 0 (control), 24, 96, and 144 hours. Mean plasma membrane fluorescence intensity (FI or IF) from a representative pixel per cell was measured by the ImageJ program. The average of the mean fluorescence intensity (FI) and its standard deviation (SD) from 50 individual cells are shown. The Pearson correlations between ABCG2 and SSEA-1 are shown in the right panels. Abbreviations: CE, coefficient; hr, hour(s).
    Fluorescence Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 45629 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 45629 article reviews
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    Image Search Results


    (a) Immunofluorescence staining of the autophagosomal protein LC3 (green fluorescence) and of the lysosomal protein LAMP1 (red fluorescence) and (b) of BECLIN 1 (green fluorescence) and Golgi-associated Golgin97 (red fluorescence) in SKOV3 cells subjected to arginine starvation. Nuclei were labelled with DAPI. Images were captured with ZEISS fluorescence microscope (Axio Imager A1) equipped with Axio Vision Software v. 4.6.3. Magnification 1000x.

    Journal: BioMed Research International

    Article Title: Single Amino Acid Arginine Deprivation Triggers Prosurvival Autophagic Response in Ovarian Carcinoma SKOV3

    doi: 10.1155/2014/505041

    Figure Lengend Snippet: (a) Immunofluorescence staining of the autophagosomal protein LC3 (green fluorescence) and of the lysosomal protein LAMP1 (red fluorescence) and (b) of BECLIN 1 (green fluorescence) and Golgi-associated Golgin97 (red fluorescence) in SKOV3 cells subjected to arginine starvation. Nuclei were labelled with DAPI. Images were captured with ZEISS fluorescence microscope (Axio Imager A1) equipped with Axio Vision Software v. 4.6.3. Magnification 1000x.

    Article Snippet: Coverslips were mounted on microscope slides and monitored under a ZEISS fluorescence microscope (Axio Imager A1) equipped with Axio Vision Software (v. 4.6.3).

    Techniques: Immunofluorescence, Staining, Fluorescence, Microscopy, Software

    Fluorescence microscopy of digestive tracts extracted from insects infected with GFP-tagged parasites. The images show the presence of green fluorescent parasites colonizing the AM (A-H), PM (I-L) and H (M-R). (A-C) Fluorescence images of the AM showing free-swimming parasites at days 1, 2 and 7 p.i., respectively. (D-H) Fluorescence and DIC images showing parasite aggregates, at day 1 p.i. (D, E), and day 4 p.i. (F-H). The asterisks indicate aggregate of parasites associated to gut wall. (I-K) Fluorescence images of the PM at days 1, 2 and 7 p.i.. (L) Fluorescence image at day 4 p.i.. The asterisk indicates a cluster of parasites in the lumen of the PM. Fluorescence and DIC images of the H at days 4 (M, P), 7 (N, Q) and 21 (O, R) p.i., respectively. Scale bars: (A-H) 50 μm; (I-K) 50 μm; (L) 10 μm; (M-R) 50 μm. A-E, I-K, M-Q and F-H and L images were performed, respectively, using a Zeiss Axioplan 2 fluorescence microscope and a Zeiss Axioplan 2 fluorescence microscope.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Monitoring of the Parasite Load in the Digestive Tract of Rhodnius prolixus by Combined qPCR Analysis and Imaging Techniques Provides New Insights into the Trypanosome Life Cycle

    doi: 10.1371/journal.pntd.0004186

    Figure Lengend Snippet: Fluorescence microscopy of digestive tracts extracted from insects infected with GFP-tagged parasites. The images show the presence of green fluorescent parasites colonizing the AM (A-H), PM (I-L) and H (M-R). (A-C) Fluorescence images of the AM showing free-swimming parasites at days 1, 2 and 7 p.i., respectively. (D-H) Fluorescence and DIC images showing parasite aggregates, at day 1 p.i. (D, E), and day 4 p.i. (F-H). The asterisks indicate aggregate of parasites associated to gut wall. (I-K) Fluorescence images of the PM at days 1, 2 and 7 p.i.. (L) Fluorescence image at day 4 p.i.. The asterisk indicates a cluster of parasites in the lumen of the PM. Fluorescence and DIC images of the H at days 4 (M, P), 7 (N, Q) and 21 (O, R) p.i., respectively. Scale bars: (A-H) 50 μm; (I-K) 50 μm; (L) 10 μm; (M-R) 50 μm. A-E, I-K, M-Q and F-H and L images were performed, respectively, using a Zeiss Axioplan 2 fluorescence microscope and a Zeiss Axioplan 2 fluorescence microscope.

    Article Snippet: Where indicated, color fluorescence image acquisition was performed using a Zeiss Axioplan 2 fluorescence microscope with a 100 W mercury lamp filtered by a Zeiss-09 filter set (excitation BP 450–490; beam splitter FT 510; emission LP 515) and acquired by a 10–24 mm Fujicolor Superia X-Tra film 400 ASA exposition.

    Techniques: Fluorescence, Microscopy, Infection

    Integrin-dependent N1E-115 cell adherence and neurite outgrowth when cells are grown on laminin. (a) The number of cells possessing neurites of at least the length of one cell body or exhibiting a flattened morphology was assessed after plating on the different matrices or following pretreatment with anti-β-1 integrin antibody. Cells were stained with rhodamine-conjugated phalloidin. Numbers are shown as a percentage of each set of 50 cells counted possessing either a neurite or a spread cell body. At least 200 cells were randomly examined, and standard deviations are indicated as vertical bars. (b) The number of cells adhering to a glass chamber slide which had been coated with different extracellular matrices was analyzed by fixing the cells 24 h following plating and staining with rhodamine-conjugated phalloidin. The number of cells per randomly selected field of view was determined on a Zeiss Axioplan fluorescence microscope, and results were compared. At least 20 fields of view were counted for each slide, and the experiments were repeated three times. Cell numbers are shown as the mean number of cells per field of view plus standard deviations shown as vertical bars.

    Journal: Molecular and Cellular Biology

    Article Title: Phosphatidylinositol 3-Kinase, Cdc42, and Rac1 Act Downstream of Ras in Integrin-Dependent Neurite Outgrowth in N1E-115 Neuroblastoma Cells

    doi:

    Figure Lengend Snippet: Integrin-dependent N1E-115 cell adherence and neurite outgrowth when cells are grown on laminin. (a) The number of cells possessing neurites of at least the length of one cell body or exhibiting a flattened morphology was assessed after plating on the different matrices or following pretreatment with anti-β-1 integrin antibody. Cells were stained with rhodamine-conjugated phalloidin. Numbers are shown as a percentage of each set of 50 cells counted possessing either a neurite or a spread cell body. At least 200 cells were randomly examined, and standard deviations are indicated as vertical bars. (b) The number of cells adhering to a glass chamber slide which had been coated with different extracellular matrices was analyzed by fixing the cells 24 h following plating and staining with rhodamine-conjugated phalloidin. The number of cells per randomly selected field of view was determined on a Zeiss Axioplan fluorescence microscope, and results were compared. At least 20 fields of view were counted for each slide, and the experiments were repeated three times. Cell numbers are shown as the mean number of cells per field of view plus standard deviations shown as vertical bars.

    Article Snippet: Slides were mounted in Mowiol–2.5% DABCO [1,4-diazabicyclo(2,2,2)octane] in a 2.5% (wt/vol) glycerol solution–0.1 M Tris (pH 8.5) (Aldrich) and viewed under a Zeiss Axioplan fluorescence microscope.

    Techniques: Staining, Fluorescence, Microscopy

    Subcellular localization of the mammalian COP1 (A) Detection of endogenous HsCOP1 protein by immunoprecipitation and immunoblot. Whole cell lysates of HeLa cells were immunoprecipitated with purified anti-COP1 antibodies from rabbit 1 (R1) or preimmune serum (Pre) as indicated, separated by SDS-PAGE, and then immunoblotted with anti-COP1 antibodies from rabbit 1, 2, and 3. The arrowheads mark the endogenous HsCOP1 protein and IgG. R1, blot probed with purified COP1 antibodies from rabbit 1; R2, blot probed purified COP1 antibodies from rabbit 2; R3, blot probed with purified COP1 antibodies from rabbit 3; Pre, preimmune serum was used for IP.(B) Localization study of endogenous HsCOP1 by subcellular fractionation. Subcellular fractions of HeLa cells were either immunoprecipitated with anti-COP1 antibodies first, or were directly separated (for all other control proteins) by SDS-PAGE and immunoblotted with antibodies against COP1, Hsp110, Hsp90, Lamin A/C, nucleoporins (mAb414), CSN2 and CSN6 from the top to the bottom panels as indicated. H, homogenate; C, cytosol; MI, mitochondria; MS, microsomes; N, nuclei; NP, nucleoplasm; NE, nuclear envelopes. (C) GFP fluorescence analysis of the subcellular localization of GFP-tagged MmCOP1 protein. N-terminal GFP-tagged MmCOP1 expression construct was transfected into COS7 cells. The cells were fixed with 4% paraformaldehyde 24 hours after transfection. Type I to III are representative of three distinct types of localization patterns, while +Triton indicates cell treated with 1% Triton in PBS for 10 min on ice and then washed three times with PBS before fixation. The subcellular localization of GFP-COP1 was observed under Zeiss Axiophot fluorescence microscope (top). The nuclei were stained by 1% DAPI in anti-fade reagent (middle). The merged pictures of the DAPI and GFP fluorescence images are shown at bottom. The same magnification was used for all the images and the scale bar for 10 micrometer was indicated within the top left image.

    Journal: BMC Cell Biology

    Article Title: An initial biochemical and cell biological characterization of the mammalian homologue of a central plant developmental switch, COP1

    doi: 10.1186/1471-2121-3-30

    Figure Lengend Snippet: Subcellular localization of the mammalian COP1 (A) Detection of endogenous HsCOP1 protein by immunoprecipitation and immunoblot. Whole cell lysates of HeLa cells were immunoprecipitated with purified anti-COP1 antibodies from rabbit 1 (R1) or preimmune serum (Pre) as indicated, separated by SDS-PAGE, and then immunoblotted with anti-COP1 antibodies from rabbit 1, 2, and 3. The arrowheads mark the endogenous HsCOP1 protein and IgG. R1, blot probed with purified COP1 antibodies from rabbit 1; R2, blot probed purified COP1 antibodies from rabbit 2; R3, blot probed with purified COP1 antibodies from rabbit 3; Pre, preimmune serum was used for IP.(B) Localization study of endogenous HsCOP1 by subcellular fractionation. Subcellular fractions of HeLa cells were either immunoprecipitated with anti-COP1 antibodies first, or were directly separated (for all other control proteins) by SDS-PAGE and immunoblotted with antibodies against COP1, Hsp110, Hsp90, Lamin A/C, nucleoporins (mAb414), CSN2 and CSN6 from the top to the bottom panels as indicated. H, homogenate; C, cytosol; MI, mitochondria; MS, microsomes; N, nuclei; NP, nucleoplasm; NE, nuclear envelopes. (C) GFP fluorescence analysis of the subcellular localization of GFP-tagged MmCOP1 protein. N-terminal GFP-tagged MmCOP1 expression construct was transfected into COS7 cells. The cells were fixed with 4% paraformaldehyde 24 hours after transfection. Type I to III are representative of three distinct types of localization patterns, while +Triton indicates cell treated with 1% Triton in PBS for 10 min on ice and then washed three times with PBS before fixation. The subcellular localization of GFP-COP1 was observed under Zeiss Axiophot fluorescence microscope (top). The nuclei were stained by 1% DAPI in anti-fade reagent (middle). The merged pictures of the DAPI and GFP fluorescence images are shown at bottom. The same magnification was used for all the images and the scale bar for 10 micrometer was indicated within the top left image.

    Article Snippet: Zeiss Axiophot fluorescence microscope was used for observation.

    Techniques: Immunoprecipitation, Purification, SDS Page, Fractionation, Mass Spectrometry, Fluorescence, Expressing, Construct, Transfection, Microscopy, Staining

    Effect of NP-1 or acyclovir on VP-16 translocation to the host cell nucleus. Cells were mock infected (a) or inoculated with HSV-2(G) that had been preincubated for 1 h at 37°C with 0.1% acetic acid (b), 25 μg of NP-1/ml (c), or 100 μg of acyclovir/ml (d). After 4 h at 37°C, cells were fixed, permeabilized, and incubated with mouse monoclonal anti-VP16 Ab, followed by incubation with goat anti-mouse IgG conjugated to FITC. Cells were visualized using a Zeiss Axioskop fluorescence microscope. The number of cells expressing VP16 in the nucleus as a percentage of the total cells visualized per high-power field were counted in four to eight fields per experiment. The graph depicts the mean ± standard deviation obtained from three to four independent experiments; the asterisk indicates a significant reduction in VP16 translocation, by the Mann-Whitney test ( P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: NP-1, a Rabbit ?-Defensin, Prevents the Entry and Intercellular Spread of Herpes Simplex Virus Type 2

    doi: 10.1128/AAC.47.2.494-500.2003

    Figure Lengend Snippet: Effect of NP-1 or acyclovir on VP-16 translocation to the host cell nucleus. Cells were mock infected (a) or inoculated with HSV-2(G) that had been preincubated for 1 h at 37°C with 0.1% acetic acid (b), 25 μg of NP-1/ml (c), or 100 μg of acyclovir/ml (d). After 4 h at 37°C, cells were fixed, permeabilized, and incubated with mouse monoclonal anti-VP16 Ab, followed by incubation with goat anti-mouse IgG conjugated to FITC. Cells were visualized using a Zeiss Axioskop fluorescence microscope. The number of cells expressing VP16 in the nucleus as a percentage of the total cells visualized per high-power field were counted in four to eight fields per experiment. The graph depicts the mean ± standard deviation obtained from three to four independent experiments; the asterisk indicates a significant reduction in VP16 translocation, by the Mann-Whitney test ( P

    Article Snippet: Cells were visualized under a Zeiss Axioskop fluorescence microscope.

    Techniques: Translocation Assay, Infection, Incubation, Fluorescence, Microscopy, Expressing, Standard Deviation, MANN-WHITNEY

    MPT0B098 inhibits hypoxia-induced F-actin rearrangement and FAK phosphorylation. a Immunofluorescent analysis of F-actin. OEC-M1 cells were treated with various concentrations, indicated as fold of IC 50 , of MPT0B098 for 18 h under hypoxic conditions, stained with phalloidin to label F-actin (red), counterstained with DAPI (blue), and then observed using an OLYMPUS florescence microscope. b Effect of MPT0B098 on FAK phosphorylation and expression. OEC-M1 cells were treated with various concentrations of MPT0B098 for 18 h.At the end of the drug treatment, cell lysates were prepared and analyzed by SDS-PAGE and Western blot. β-Actin was used as an internal control. c Each bar depicts the mean of the relative intensities of Twist and SNAI2/Slug from three independent experiments (* p

    Journal: Journal of Biomedical Science

    Article Title: Novel microtubule inhibitor MPT0B098 inhibits hypoxia-induced epithelial-to-mesenchymal transition in head and neck squamous cell carcinoma

    doi: 10.1186/s12929-018-0432-6

    Figure Lengend Snippet: MPT0B098 inhibits hypoxia-induced F-actin rearrangement and FAK phosphorylation. a Immunofluorescent analysis of F-actin. OEC-M1 cells were treated with various concentrations, indicated as fold of IC 50 , of MPT0B098 for 18 h under hypoxic conditions, stained with phalloidin to label F-actin (red), counterstained with DAPI (blue), and then observed using an OLYMPUS florescence microscope. b Effect of MPT0B098 on FAK phosphorylation and expression. OEC-M1 cells were treated with various concentrations of MPT0B098 for 18 h.At the end of the drug treatment, cell lysates were prepared and analyzed by SDS-PAGE and Western blot. β-Actin was used as an internal control. c Each bar depicts the mean of the relative intensities of Twist and SNAI2/Slug from three independent experiments (* p

    Article Snippet: Cells were observed with an OLYMPUS fluorescence microscope.

    Techniques: Staining, Microscopy, Expressing, SDS Page, Western Blot

    Binding of FITC-labeled Vg to macrophages. Macrophage suspensions and FITC-labeled Vg were incubated together at room temperature for 30 min, and then 60 µl of cell suspensions was sampled to make smears for microscopic examination under an Olympus BX51 fluorescence microscope. For control, red blood cells were used, and treated similarly. A and C are the images under a fluorescent field, and B and D are the images under a bright light field. (A and B) Macrophages. (C and D) Red blood cells. Scale bars: 10 µm.

    Journal: PLoS ONE

    Article Title: Vitellogenin Functions as a Multivalent Pattern Recognition Receptor with an Opsonic Activity

    doi: 10.1371/journal.pone.0001940

    Figure Lengend Snippet: Binding of FITC-labeled Vg to macrophages. Macrophage suspensions and FITC-labeled Vg were incubated together at room temperature for 30 min, and then 60 µl of cell suspensions was sampled to make smears for microscopic examination under an Olympus BX51 fluorescence microscope. For control, red blood cells were used, and treated similarly. A and C are the images under a fluorescent field, and B and D are the images under a bright light field. (A and B) Macrophages. (C and D) Red blood cells. Scale bars: 10 µm.

    Article Snippet: Aliquots of 10 µl of the microbial suspensions were applied to microscope slides, and binding of FITC-labeled Vg to the microbial cells was observed under an Olympus BX51 fluorescence microscope.

    Techniques: Binding Assay, Labeling, Incubation, Fluorescence, Microscopy

    Binding of FITC-labeled Vg to microbial cells. Vg was labeled by FITC, and FITC-labeled Vg was incubated with Gram-negative bacterium E. coli , Gram-positive bacterium S. aureus and fungus P. pastoris , respectively. After washing three times with 25 mM Tris-HCl buffer containing 137 mM NaCl and 3 mM KCl (TBS; pH 7.6), the microbes were harvested by centrifugation, applied to microscope slides, and observed under an Olympus BX51 fluorescence microscope. The microbes treated with FITC-labeled BSA and FITC-labeled apoB were processed under the same conditions. (A, D and G) Binding of FITC-labeled Vg to E. coli, S. aureus and P. pastoris ; (B, E and H) Non-binding of FITC-labeled BSA to E. coli, S. aureus and P. pastoris ; (C, F and I) Non-binding of FITC-labeled apoB to E. coli, S. aureus and P. pastoris . A, D and G are the images under a fluorescent field, while B, E, H, C, F and H are the merged images under fluorescent and bright field channels. Scale bars: 20 µm.

    Journal: PLoS ONE

    Article Title: Vitellogenin Functions as a Multivalent Pattern Recognition Receptor with an Opsonic Activity

    doi: 10.1371/journal.pone.0001940

    Figure Lengend Snippet: Binding of FITC-labeled Vg to microbial cells. Vg was labeled by FITC, and FITC-labeled Vg was incubated with Gram-negative bacterium E. coli , Gram-positive bacterium S. aureus and fungus P. pastoris , respectively. After washing three times with 25 mM Tris-HCl buffer containing 137 mM NaCl and 3 mM KCl (TBS; pH 7.6), the microbes were harvested by centrifugation, applied to microscope slides, and observed under an Olympus BX51 fluorescence microscope. The microbes treated with FITC-labeled BSA and FITC-labeled apoB were processed under the same conditions. (A, D and G) Binding of FITC-labeled Vg to E. coli, S. aureus and P. pastoris ; (B, E and H) Non-binding of FITC-labeled BSA to E. coli, S. aureus and P. pastoris ; (C, F and I) Non-binding of FITC-labeled apoB to E. coli, S. aureus and P. pastoris . A, D and G are the images under a fluorescent field, while B, E, H, C, F and H are the merged images under fluorescent and bright field channels. Scale bars: 20 µm.

    Article Snippet: Aliquots of 10 µl of the microbial suspensions were applied to microscope slides, and binding of FITC-labeled Vg to the microbial cells was observed under an Olympus BX51 fluorescence microscope.

    Techniques: Binding Assay, Labeling, Incubation, Centrifugation, Microscopy, Fluorescence

    Effects of interleukin (IL)-17A and IL-22 on the expression of stem-cell markers in passage-one epidermal sheets. The epidermal sheet was prepared as described in Materials and methods. (A) After 3–5 days of cultivation, passage-one epidermal cells had grown from the edge of the skin explant to form a tongue-like epidermal sheet. The dotted line indicates the edge of the skin explant. Arrows indicate the leading-edge zone of the epidermal sheet. The epidermal sheet was treated with recombinant IL-17A (10 ng/ml) and IL-22 (10 ng/ml) for 24 h. Shifts in the expression profiles of (B) K15 and (C) integrin β1 (ITGB1) in the epidermal sheet were examined by immunofluorescence staining. The slides were counterstained with DAPI to identify nuclei. Immunostaining was observed using an Olympus IX71 fluorescence microscope. Scale bar, 50 µ m.

    Journal: International Journal of Molecular Medicine

    Article Title: Asymmetric stem-cell division ensures sustained keratinocyte hyperproliferation in psoriatic skin lesions

    doi: 10.3892/ijmm.2015.2445

    Figure Lengend Snippet: Effects of interleukin (IL)-17A and IL-22 on the expression of stem-cell markers in passage-one epidermal sheets. The epidermal sheet was prepared as described in Materials and methods. (A) After 3–5 days of cultivation, passage-one epidermal cells had grown from the edge of the skin explant to form a tongue-like epidermal sheet. The dotted line indicates the edge of the skin explant. Arrows indicate the leading-edge zone of the epidermal sheet. The epidermal sheet was treated with recombinant IL-17A (10 ng/ml) and IL-22 (10 ng/ml) for 24 h. Shifts in the expression profiles of (B) K15 and (C) integrin β1 (ITGB1) in the epidermal sheet were examined by immunofluorescence staining. The slides were counterstained with DAPI to identify nuclei. Immunostaining was observed using an Olympus IX71 fluorescence microscope. Scale bar, 50 µ m.

    Article Snippet: Immunostaining was subsequently observed using an Olympus IX71 fluorescence microscope.

    Techniques: Expressing, Recombinant, Immunofluorescence, Staining, Immunostaining, Fluorescence, Microscopy

    In vitro analyses for template-DNA strand co-segregation in trypsin-dissociated psoriatic keratinocytes using BrdU pulse-chase labeling. Primary keratinocytes (passage 2) were selectively cultured from psoriatic plaques and from normal skin tissues, and were then plated singly on collagen-coated coverslips in 6-well tissue culture plates. After the cells had attached, 10 µ M BrdU was added for a 30 h pulse-labeling (the doubling time for keratinocytes). The cells were washed and re-fed with fresh medium containing 5 µ M cytochalasin D for an additional 30 h to arrest cytokinesis. After the chase period, the cells were fixed for BrdU immunostaining. The slides were counterstained with DAPI to identify nuclei. Immunostaining was observed using an Olympus IX71 fluorescence microscope. (A) Cell pairs were visualized for BrdU (green) and DAPI (blue). Depicted is a representative image of an immunostained pair of cells: the top row shows a cell pair with symmetric inheritance of BrdU-labeled DNA, and the bottom row shows an asymmetric pair. The dotted lines in the top and bottom rows delineate one cell pair each. The areas boxed in white in the low-magnification images (the far left panels) indicate the locations of the cell pairs in the slide. The percentage of cells presenting asymmetric inheritance of template DNA strands is shown in (B). Data represent the means ± SD from 3 independent experiments. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Asymmetric stem-cell division ensures sustained keratinocyte hyperproliferation in psoriatic skin lesions

    doi: 10.3892/ijmm.2015.2445

    Figure Lengend Snippet: In vitro analyses for template-DNA strand co-segregation in trypsin-dissociated psoriatic keratinocytes using BrdU pulse-chase labeling. Primary keratinocytes (passage 2) were selectively cultured from psoriatic plaques and from normal skin tissues, and were then plated singly on collagen-coated coverslips in 6-well tissue culture plates. After the cells had attached, 10 µ M BrdU was added for a 30 h pulse-labeling (the doubling time for keratinocytes). The cells were washed and re-fed with fresh medium containing 5 µ M cytochalasin D for an additional 30 h to arrest cytokinesis. After the chase period, the cells were fixed for BrdU immunostaining. The slides were counterstained with DAPI to identify nuclei. Immunostaining was observed using an Olympus IX71 fluorescence microscope. (A) Cell pairs were visualized for BrdU (green) and DAPI (blue). Depicted is a representative image of an immunostained pair of cells: the top row shows a cell pair with symmetric inheritance of BrdU-labeled DNA, and the bottom row shows an asymmetric pair. The dotted lines in the top and bottom rows delineate one cell pair each. The areas boxed in white in the low-magnification images (the far left panels) indicate the locations of the cell pairs in the slide. The percentage of cells presenting asymmetric inheritance of template DNA strands is shown in (B). Data represent the means ± SD from 3 independent experiments. * P

    Article Snippet: Immunostaining was subsequently observed using an Olympus IX71 fluorescence microscope.

    Techniques: In Vitro, Pulse Chase, Labeling, Cell Culture, Immunostaining, Fluorescence, Microscopy

    Determination of the transition from stem cells to transit-amplifying (TA) cells in BrdU pulse-chased anaphase cells using dual immunostaining for BrdU segregation and K15 expression. (A) Primary keratinocytes were cultured and pulse-chased with BrdU as described in Fig. 3 . Cell pairs were co-immunostained for BrdU and K15. Depicted is a representative pair of cells demonstrating K15 expression and symmetric BrdU staining (top row), one daughter cell expressing K15 and symmetric BrdU staining (middle row), and one daughter cell expressing K15 and asymmetric BrdU staining (bottom row). The slides were counterstained with Hoechst 33258 to identify nuclei. Immunostaining was observed using an Olympus IX71 fluorescence microscope. Representative anaphase cells are shown in differential interference contrast (DIC) (furthest right panel), melanin pigment (brownish) was also visualized in the cells, which potentially affects the nuclear staining by Hoechst 33258 dye. Note that DNA template strands (BrdU-unlabeled) segregate to the less-differentiated progeny (K15-positive cells). The dotted lines delineate one cell pair each. Scale bar, 10 µ m. (B) The percentage of cell pairs presenting asymmetric inheritance of template DNA strands and K15 expression. Data represent the means ± SD from 3 independent experiments. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Asymmetric stem-cell division ensures sustained keratinocyte hyperproliferation in psoriatic skin lesions

    doi: 10.3892/ijmm.2015.2445

    Figure Lengend Snippet: Determination of the transition from stem cells to transit-amplifying (TA) cells in BrdU pulse-chased anaphase cells using dual immunostaining for BrdU segregation and K15 expression. (A) Primary keratinocytes were cultured and pulse-chased with BrdU as described in Fig. 3 . Cell pairs were co-immunostained for BrdU and K15. Depicted is a representative pair of cells demonstrating K15 expression and symmetric BrdU staining (top row), one daughter cell expressing K15 and symmetric BrdU staining (middle row), and one daughter cell expressing K15 and asymmetric BrdU staining (bottom row). The slides were counterstained with Hoechst 33258 to identify nuclei. Immunostaining was observed using an Olympus IX71 fluorescence microscope. Representative anaphase cells are shown in differential interference contrast (DIC) (furthest right panel), melanin pigment (brownish) was also visualized in the cells, which potentially affects the nuclear staining by Hoechst 33258 dye. Note that DNA template strands (BrdU-unlabeled) segregate to the less-differentiated progeny (K15-positive cells). The dotted lines delineate one cell pair each. Scale bar, 10 µ m. (B) The percentage of cell pairs presenting asymmetric inheritance of template DNA strands and K15 expression. Data represent the means ± SD from 3 independent experiments. * P

    Article Snippet: Immunostaining was subsequently observed using an Olympus IX71 fluorescence microscope.

    Techniques: Immunostaining, Expressing, Cell Culture, BrdU Staining, Fluorescence, Microscopy, Staining

    Genes encoding pellicular proteins are AP2-O targets. A. The pellicle structure and its components are illustrated. Targets are highlighted in red. In addition to the proteins mentioned in the text, targets contained several genes encoding putative pellicular/IMC proteins, viz. apicortin, tubulin-tyrosine ligase (TTL), and small heat shock-related protein 20 (HSP20). Apicortin is involved in the stabilization of microtubules [ 59 ]. TTL, an enzyme adding a tyrosine to the carboxyl end of tubulin, marks the plus ends of growing microtubules and regulates microtubule growth at this site [ 60 ]. HSP20 is involved in ookinete motility [ 51 , 61 ]. MyoA, myosin A; ADF1, actin-depolymerizing factor 1; SPM, subpellicular microtubule. B. IMC1i was expressed as a GFP-tagged protein using a P . berghei centromere plasmid. Ookinetes were cultured in vitro and observed by fluorescence microscopy at 24 h after fertilization. The apical end of the ookinete is indicated by an arrowhead. Left, merged image of GFP and nuclear staining with Hoechst 33342. Right, optical transmission image. Scale bar, 10 μm. C. Giemsa-stained image of wild-type (left) and IMC1i (-) (right) ookinetes, 24 h after fertilization. Scale bar, 10 μm. D. Fluorescence microscopy image of ookinetes expressing GFP-tagged PUA26. Left, merged image of GFP and nuclear staining with Hoechst 33342. Right, optical transmission image. Scale bar, 10 μm. E. Immunoelectron microscopy image. Cross-sections of two ookinetes expressing GFP-tagged PUA26 are shown. Immunoelectron microscopy was performed with anti-GFP antibodies. Colloidal gold particles (15 nm) are mainly localized at the IMC, which is the layer of high-electron density beneath the plasma membrane. Subpellicular microtubules are indicated by arrowheads. Scale bar, 1 μm. F. Giemsa-stained image of wild-type (left) and PUA26 (-) (right) ookinetes at 24 h after fertilization. Scale bar, 10 μm. G. Height—width ratios were compared between wild-type and PUA26 (-) ookinetes. Ookinetes were cultured for 24 h and stained with Giemsa. Micrographs were obtained with an Olympus BX60 fluorescence microscope. Height—width ratios of ookinetes were measured with the AquaCosmos software (Hamamatsu Photonic System). In total, 100 ookinetes were analyzed in each parasite population. Bars represent mean ± SE. H. The numbers of parasites associated with the midgut were compared between wild-type and PUA26 (-) parasites at 24 h after an infective blood meal by mosquitoes. Data are the mean ± SE of three independent experiments using 20 mosquitoes each. Only parasites on a single side of the midgut were counted.

    Journal: PLoS Pathogens

    Article Title: Genome-Wide Identification of the Target Genes of AP2-O, a Plasmodium AP2-Family Transcription Factor

    doi: 10.1371/journal.ppat.1004905

    Figure Lengend Snippet: Genes encoding pellicular proteins are AP2-O targets. A. The pellicle structure and its components are illustrated. Targets are highlighted in red. In addition to the proteins mentioned in the text, targets contained several genes encoding putative pellicular/IMC proteins, viz. apicortin, tubulin-tyrosine ligase (TTL), and small heat shock-related protein 20 (HSP20). Apicortin is involved in the stabilization of microtubules [ 59 ]. TTL, an enzyme adding a tyrosine to the carboxyl end of tubulin, marks the plus ends of growing microtubules and regulates microtubule growth at this site [ 60 ]. HSP20 is involved in ookinete motility [ 51 , 61 ]. MyoA, myosin A; ADF1, actin-depolymerizing factor 1; SPM, subpellicular microtubule. B. IMC1i was expressed as a GFP-tagged protein using a P . berghei centromere plasmid. Ookinetes were cultured in vitro and observed by fluorescence microscopy at 24 h after fertilization. The apical end of the ookinete is indicated by an arrowhead. Left, merged image of GFP and nuclear staining with Hoechst 33342. Right, optical transmission image. Scale bar, 10 μm. C. Giemsa-stained image of wild-type (left) and IMC1i (-) (right) ookinetes, 24 h after fertilization. Scale bar, 10 μm. D. Fluorescence microscopy image of ookinetes expressing GFP-tagged PUA26. Left, merged image of GFP and nuclear staining with Hoechst 33342. Right, optical transmission image. Scale bar, 10 μm. E. Immunoelectron microscopy image. Cross-sections of two ookinetes expressing GFP-tagged PUA26 are shown. Immunoelectron microscopy was performed with anti-GFP antibodies. Colloidal gold particles (15 nm) are mainly localized at the IMC, which is the layer of high-electron density beneath the plasma membrane. Subpellicular microtubules are indicated by arrowheads. Scale bar, 1 μm. F. Giemsa-stained image of wild-type (left) and PUA26 (-) (right) ookinetes at 24 h after fertilization. Scale bar, 10 μm. G. Height—width ratios were compared between wild-type and PUA26 (-) ookinetes. Ookinetes were cultured for 24 h and stained with Giemsa. Micrographs were obtained with an Olympus BX60 fluorescence microscope. Height—width ratios of ookinetes were measured with the AquaCosmos software (Hamamatsu Photonic System). In total, 100 ookinetes were analyzed in each parasite population. Bars represent mean ± SE. H. The numbers of parasites associated with the midgut were compared between wild-type and PUA26 (-) parasites at 24 h after an infective blood meal by mosquitoes. Data are the mean ± SE of three independent experiments using 20 mosquitoes each. Only parasites on a single side of the midgut were counted.

    Article Snippet: Micrographs were obtained with an Olympus BX60 fluorescence microscope.

    Techniques: Plasmid Preparation, Cell Culture, In Vitro, Fluorescence, Microscopy, Staining, Transmission Assay, Expressing, Immuno-Electron Microscopy, Software

    Higher expression of miR-351 shifts the FUdR-induced cell death morphologies from necrosis to apoptosis. (A) At 48 hours after transfection with the vehicle, non-silencing siRNA (NSsi) or the mature miR-351-5p mimic (miR351m), the F28-7 cells were treated with or without 1 μM FUdR for 21 h, and then stained with DAPI. Morphological changes were analyzed by Olympus BX61 fluorescence microscope at 400× magnification. (B) Percentage of necrosis, apoptosis, and normal cell morphologies in FUdR-treated F28-7 cells transfected with NSsi or miR351m. (C) At 48 hours after transfection with the vehicle, non-silencing siRNA (NSsi) or the miR-351 mimic (miR351m), the F28-7 cells were treated with or without 1 μM FUdR for 21 h, and analyzed by western blotting for anti-HMGB1 antibody. As a negative control, F28-7-A cells were treated with or without 1 μM FUdR for 21 h, and examined by western blotting. Two additional independent experiments gave similar results.

    Journal: PLoS ONE

    Article Title: MicroRNA-351 Regulates Two-Types of Cell Death, Necrosis and Apoptosis, Induced by 5-fluoro-2'-deoxyuridine

    doi: 10.1371/journal.pone.0153130

    Figure Lengend Snippet: Higher expression of miR-351 shifts the FUdR-induced cell death morphologies from necrosis to apoptosis. (A) At 48 hours after transfection with the vehicle, non-silencing siRNA (NSsi) or the mature miR-351-5p mimic (miR351m), the F28-7 cells were treated with or without 1 μM FUdR for 21 h, and then stained with DAPI. Morphological changes were analyzed by Olympus BX61 fluorescence microscope at 400× magnification. (B) Percentage of necrosis, apoptosis, and normal cell morphologies in FUdR-treated F28-7 cells transfected with NSsi or miR351m. (C) At 48 hours after transfection with the vehicle, non-silencing siRNA (NSsi) or the miR-351 mimic (miR351m), the F28-7 cells were treated with or without 1 μM FUdR for 21 h, and analyzed by western blotting for anti-HMGB1 antibody. As a negative control, F28-7-A cells were treated with or without 1 μM FUdR for 21 h, and examined by western blotting. Two additional independent experiments gave similar results.

    Article Snippet: Morphological changes were analyzed by Olympus BX61 fluorescence microscope at 400× magnification. (PDF) Click here for additional data file.

    Techniques: Expressing, Transfection, Staining, Fluorescence, Microscopy, Western Blot, Negative Control

    The impact of CuO NMs and CuSO 4 on Caco-2 cell morphology. a ) Following differentiation, cell monolayers were exposed to cell culture medium (control) or 6.34 μg/cm 2 Cu of CuO NMs and CuSO 4 for 24 h and then fixed and stained for the tight junction protein ZO-1 (green) and nucleus with DAPI (blue). The images were obtained with Zeiss LSM880 confocal microscope using the Zen program for data analyses. Scale bar = 20 μm. b ) Assessment of cell morphology using SEM. Differentiated and undifferentiated Caco-2 cells were exposed to 12.68 μg/cm 2 Cu of CuO NMs for 24 h and then were washed, fixed, dehydrated, dried and examined by FIB/SEM. i and ii are control differentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. iii and iv are control undifferentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. v and vi are differentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively and vii and viii are undifferentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively

    Journal: Particle and Fibre Toxicology

    Article Title: Impact of copper oxide nanomaterials on differentiated and undifferentiated Caco-2 intestinal epithelial cells; assessment of cytotoxicity, barrier integrity, cytokine production and nanomaterial penetration

    doi: 10.1186/s12989-017-0211-7

    Figure Lengend Snippet: The impact of CuO NMs and CuSO 4 on Caco-2 cell morphology. a ) Following differentiation, cell monolayers were exposed to cell culture medium (control) or 6.34 μg/cm 2 Cu of CuO NMs and CuSO 4 for 24 h and then fixed and stained for the tight junction protein ZO-1 (green) and nucleus with DAPI (blue). The images were obtained with Zeiss LSM880 confocal microscope using the Zen program for data analyses. Scale bar = 20 μm. b ) Assessment of cell morphology using SEM. Differentiated and undifferentiated Caco-2 cells were exposed to 12.68 μg/cm 2 Cu of CuO NMs for 24 h and then were washed, fixed, dehydrated, dried and examined by FIB/SEM. i and ii are control differentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. iii and iv are control undifferentiated Caco-2 cells enlarged X 5000 and X 10000 respectively. v and vi are differentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively and vii and viii are undifferentiated Caco-2 cells exposed to 12.68 μg/cm 2 Cu of CuO NMs enlarged X 5000 and X 10000 respectively

    Article Snippet: Cells were visualized using Zeiss LSM880 confocal microscope for ZO-1 or Zeiss fluorescent Microscope, Carl Zeiss Axio Scope A 1 Upright Research Microscope for nuclear counting (Germany) and the results analysed using the Zen program and Image J software.

    Techniques: Cell Culture, Staining, Microscopy

    Indirect immunofluorescence analysis of surface ABCG2, SSEA-1, and SSEA-4 coexpression during differentiation. (A): WA01 cells (control); (B and C): WA01 cells treated with 50 ng/mL or 100 ng/mL of recombinant BMP-4 for 6 days. C1 indicates BMP-4 concentration 1 (i.e., 50 ng/mL) and C2 denotes BMP-4 concentration 2 (i.e., 100 ng/mL). The images were acquired with an Axiovert 200 fluorescence microscope (Zeiss). Bar, 50 μm. (D and E): Quantitative analysis of the coexpression of surface ABCG2 and SSEA-1 in WA01 cells (D) and WA09 cells (E) under the treatment of the cells with 100 ng/mL of BMP-4 for 0 (control), 24, 96, and 144 hours. Mean plasma membrane fluorescence intensity (FI or IF) from a representative pixel per cell was measured by the ImageJ program. The average of the mean fluorescence intensity (FI) and its standard deviation (SD) from 50 individual cells are shown. The Pearson correlations between ABCG2 and SSEA-1 are shown in the right panels. Abbreviations: CE, coefficient; hr, hour(s).

    Journal: Stem cells (Dayton, Ohio)

    Article Title: Regulation and Expression of the ATP-binding Cassette Transporter ABCG2 in Human Embryonic Stem Cells

    doi: 10.1002/stem.1195

    Figure Lengend Snippet: Indirect immunofluorescence analysis of surface ABCG2, SSEA-1, and SSEA-4 coexpression during differentiation. (A): WA01 cells (control); (B and C): WA01 cells treated with 50 ng/mL or 100 ng/mL of recombinant BMP-4 for 6 days. C1 indicates BMP-4 concentration 1 (i.e., 50 ng/mL) and C2 denotes BMP-4 concentration 2 (i.e., 100 ng/mL). The images were acquired with an Axiovert 200 fluorescence microscope (Zeiss). Bar, 50 μm. (D and E): Quantitative analysis of the coexpression of surface ABCG2 and SSEA-1 in WA01 cells (D) and WA09 cells (E) under the treatment of the cells with 100 ng/mL of BMP-4 for 0 (control), 24, 96, and 144 hours. Mean plasma membrane fluorescence intensity (FI or IF) from a representative pixel per cell was measured by the ImageJ program. The average of the mean fluorescence intensity (FI) and its standard deviation (SD) from 50 individual cells are shown. The Pearson correlations between ABCG2 and SSEA-1 are shown in the right panels. Abbreviations: CE, coefficient; hr, hour(s).

    Article Snippet: The images were acquired under an Axiovert 200 fluorescence microscope (Zeiss).

    Techniques: Immunofluorescence, Recombinant, Concentration Assay, Fluorescence, Microscopy, Standard Deviation