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  • 99
    Millipore hoechst 33258
    The leucine-rich NES of Sox10 mediates nucleocytoplasmic shuttling of Sox10. GFP-Sox10 and GFP-Sox10ΔNES were transfected into HeLa cells and analyzed in heterokaryon assays. (A) The wild-type GFP-Sox10 fusion protein was rapidly translocated from the HeLa donor nucleus into the surrounding 3T3 acceptor nuclei. Under LMB treatment, GFP-Sox10 was not able to shuttle into the acceptor nuclei (C and H) in contrast to pUL96 (G). (E) The introduction of the leucine to alanine mutations resulted in a strong inhibition of nucleocytoplasmic transport of GFP-Sox10ΔNES fusion protein. (B, D, F, and I) <t>Hoechst</t> 33258 counterstains.
    Hoechst 33258, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12885 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 12885 article reviews
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    88
    Millipore bisbenzimide solution
    Immunofluorescence labeling with three different mAbs directed against rat postnatal cortical membrane preparations in the developing rat cortex. A , mab 10 ; B , mab 11 ; C , mab 942 . Top , Frontal sections from P6 cortex; bottom , frontal sections from E19 cortex. To the left of each fluorescent image is a micrograph of the left half of the same section counterstained with <t>bisbenzimide</t> to illustrate the cortical layering. MZ , Marginal zone; CP , cortical plate; SP , subplate zone; IZ ; intermediate zone; VZ , ventricular zone. Scale bars, 100 μm.
    Bisbenzimide Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 88 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    bisbenzimide solution - by Bioz Stars, 2020-05
    88/100 stars
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    Image Search Results


    The leucine-rich NES of Sox10 mediates nucleocytoplasmic shuttling of Sox10. GFP-Sox10 and GFP-Sox10ΔNES were transfected into HeLa cells and analyzed in heterokaryon assays. (A) The wild-type GFP-Sox10 fusion protein was rapidly translocated from the HeLa donor nucleus into the surrounding 3T3 acceptor nuclei. Under LMB treatment, GFP-Sox10 was not able to shuttle into the acceptor nuclei (C and H) in contrast to pUL96 (G). (E) The introduction of the leucine to alanine mutations resulted in a strong inhibition of nucleocytoplasmic transport of GFP-Sox10ΔNES fusion protein. (B, D, F, and I) Hoechst 33258 counterstains.

    Journal: Molecular and Cellular Biology

    Article Title: Sox10 Is an Active Nucleocytoplasmic Shuttle Protein, and Shuttling Is Crucial for Sox10-Mediated Transactivation

    doi: 10.1128/MCB.22.16.5826-5834.2002

    Figure Lengend Snippet: The leucine-rich NES of Sox10 mediates nucleocytoplasmic shuttling of Sox10. GFP-Sox10 and GFP-Sox10ΔNES were transfected into HeLa cells and analyzed in heterokaryon assays. (A) The wild-type GFP-Sox10 fusion protein was rapidly translocated from the HeLa donor nucleus into the surrounding 3T3 acceptor nuclei. Under LMB treatment, GFP-Sox10 was not able to shuttle into the acceptor nuclei (C and H) in contrast to pUL96 (G). (E) The introduction of the leucine to alanine mutations resulted in a strong inhibition of nucleocytoplasmic transport of GFP-Sox10ΔNES fusion protein. (B, D, F, and I) Hoechst 33258 counterstains.

    Article Snippet: After secondary antibody incubation, Hoechst 33258 (5 μg/ml; Sigma, Taufkirchen, Germany) were added for 5 min. At least 25 heterokaryons were analyzed for each experiment and nuclear shuttling was only scored positive when a minimum of 80% of the heterokaryons showed a positive staining of the investigated protein.

    Techniques: Transfection, Inhibition

    Nuclear localization of endogenous PC4, analyzed by immunofluorescence and confocal microscopy. (A) A total of 7 × 10 4 ), followed by incubation with goat anti-rabbit FITC-conjugated antibody. Nuclei were detected by Hoechst 33258 dye (corresponding photomicrographs on the right). Bar, 30 μm. (B) Percentage of cells with endogenous PC4 staining that was cytoplasmic only (black bars), nuclear and cytoplasmic (gray bars), or nuclear only (white bars). Values are calculated for each category as the percentages of the total number of cells scored at each time point (within three fields for each experiment). Means ± the SEM are from three independent experiments. The total number of cells counted for each time point is indicated at the top of the corresponding bar. (C) Confocal microscopy of endogenous PC4. A total of 7 × 10 4 C2C7 cells were seeded onto circular coverslips placed in 35-mm dishes. Cultures were treated, and endogenous PC4 was detected as in panel A. Nuclei are visualized in red by propidium iodide (Pr.i.). The panels on the left (merge) show the overlay of the green and red staining (in orange-green), which indicates the presence of endogenous PC4 protein in the nucleus. Bar, 10 μm.

    Journal: Molecular and Cellular Biology

    Article Title: PC4 Coactivates MyoD by Relieving the Histone Deacetylase 4-Mediated Inhibition of Myocyte Enhancer Factor 2C

    doi: 10.1128/MCB.25.6.2242-2259.2005

    Figure Lengend Snippet: Nuclear localization of endogenous PC4, analyzed by immunofluorescence and confocal microscopy. (A) A total of 7 × 10 4 ), followed by incubation with goat anti-rabbit FITC-conjugated antibody. Nuclei were detected by Hoechst 33258 dye (corresponding photomicrographs on the right). Bar, 30 μm. (B) Percentage of cells with endogenous PC4 staining that was cytoplasmic only (black bars), nuclear and cytoplasmic (gray bars), or nuclear only (white bars). Values are calculated for each category as the percentages of the total number of cells scored at each time point (within three fields for each experiment). Means ± the SEM are from three independent experiments. The total number of cells counted for each time point is indicated at the top of the corresponding bar. (C) Confocal microscopy of endogenous PC4. A total of 7 × 10 4 C2C7 cells were seeded onto circular coverslips placed in 35-mm dishes. Cultures were treated, and endogenous PC4 was detected as in panel A. Nuclei are visualized in red by propidium iodide (Pr.i.). The panels on the left (merge) show the overlay of the green and red staining (in orange-green), which indicates the presence of endogenous PC4 protein in the nucleus. Bar, 10 μm.

    Article Snippet: To detect nuclei, cells were incubated at the end of the immunofluorescence staining procedure for 2 min in Hoechst 33258 dye diluted in PBS at 1 μg/ml (Sigma), washed twice in PBS, and mounted as described above.

    Techniques: Immunofluorescence, Confocal Microscopy, Incubation, Staining

    Doxorubicin (DOX) and etoposide (ETO) induced nucleus swelling, while differently affecting nucleus morphology. ( A ) Schematic of nucleus swelling and nuclear envelope topography measured using a hemocytometer, confocal microscopy, and CNT/AFM probes systems. ( B ) DOX- and ETO-treated cells were harvested and the nuclei were extracted. Nuclear extracts were seeded on a hemocytometer and visually measured using phase contrast microscopy (scale bar represents 50 μm). ( C ) Nuclear extracts were seeded on coverslips for 15 min and stained with nucleus targeting dies such as Hoechst 33258 and PI dyes that were detected using confocal microscopy. Representative images shown are the merged images for staining with Hoechst 33258 (blue color) and PI (red color; scale bar represents 10 μm).

    Journal: Scientific Reports

    Article Title: Structural and functional analysis of cell adhesion and nuclear envelope nano-topography in cell death

    doi: 10.1038/srep15623

    Figure Lengend Snippet: Doxorubicin (DOX) and etoposide (ETO) induced nucleus swelling, while differently affecting nucleus morphology. ( A ) Schematic of nucleus swelling and nuclear envelope topography measured using a hemocytometer, confocal microscopy, and CNT/AFM probes systems. ( B ) DOX- and ETO-treated cells were harvested and the nuclei were extracted. Nuclear extracts were seeded on a hemocytometer and visually measured using phase contrast microscopy (scale bar represents 50 μm). ( C ) Nuclear extracts were seeded on coverslips for 15 min and stained with nucleus targeting dies such as Hoechst 33258 and PI dyes that were detected using confocal microscopy. Representative images shown are the merged images for staining with Hoechst 33258 (blue color) and PI (red color; scale bar represents 10 μm).

    Article Snippet: Hoechst 33258 reagent (5 μM; Sigma-Aldrich Co. LLC) for nuclear staining was added to the samples for 30 min at room temperature, and samples were washed with PBS for 3 times.

    Techniques: Confocal Microscopy, Microscopy, Staining

    Inhibition of pan-caspase suppressed nuclear envelope rupturing and destruction of the nuclear pore complex, and regained endonuclease G translocation in the nuclei by etoposide (ETO). ( A ) Cells treated with ETO and cells co-treated with z-VAD were harvested. Nuclear extracts were seeded on culture dishes for 15 min and the nuclei were fixed. We measured nuclear envelope topography using a CNT/AFM probes system. Images shown are representative of 3D topography at 30-, 10-, and 2-μm scales. See also Supplementary Fig. S7 . ( B ) Cells treated with ETO and co-treated with z-VAD regulated endonuclease G (green color) translocation levels, indicated by immunofluorescence staining and Hoechst 33258 (blue color) that was used for nuclear staining and were measured using confocal microscopy (scale bar represents 10 μm) at 72 h. Arrow indicates nuclear envelope ruptures (collapse Hoechst 33258 intensity) and/or endonuclease G translocation which also is also shown in Supplementary Fig. S8 . ( C ) Translocation endonuclease G levels stained by immunofluorescence and Hoechst 33258 for nuclei that were measured using the Cellomics ArrayScan HCS Reader for at least 200 cells. Endonuclease G intensity was dependent on Hoechst 33258-positive area that is shown in the histogram compared to the control (statistical analysis P-value of *P

    Journal: Scientific Reports

    Article Title: Structural and functional analysis of cell adhesion and nuclear envelope nano-topography in cell death

    doi: 10.1038/srep15623

    Figure Lengend Snippet: Inhibition of pan-caspase suppressed nuclear envelope rupturing and destruction of the nuclear pore complex, and regained endonuclease G translocation in the nuclei by etoposide (ETO). ( A ) Cells treated with ETO and cells co-treated with z-VAD were harvested. Nuclear extracts were seeded on culture dishes for 15 min and the nuclei were fixed. We measured nuclear envelope topography using a CNT/AFM probes system. Images shown are representative of 3D topography at 30-, 10-, and 2-μm scales. See also Supplementary Fig. S7 . ( B ) Cells treated with ETO and co-treated with z-VAD regulated endonuclease G (green color) translocation levels, indicated by immunofluorescence staining and Hoechst 33258 (blue color) that was used for nuclear staining and were measured using confocal microscopy (scale bar represents 10 μm) at 72 h. Arrow indicates nuclear envelope ruptures (collapse Hoechst 33258 intensity) and/or endonuclease G translocation which also is also shown in Supplementary Fig. S8 . ( C ) Translocation endonuclease G levels stained by immunofluorescence and Hoechst 33258 for nuclei that were measured using the Cellomics ArrayScan HCS Reader for at least 200 cells. Endonuclease G intensity was dependent on Hoechst 33258-positive area that is shown in the histogram compared to the control (statistical analysis P-value of *P

    Article Snippet: Hoechst 33258 reagent (5 μM; Sigma-Aldrich Co. LLC) for nuclear staining was added to the samples for 30 min at room temperature, and samples were washed with PBS for 3 times.

    Techniques: Inhibition, Translocation Assay, Immunofluorescence, Staining, Confocal Microscopy

    Doxorubicin (DOX) and etoposide (ETO) induced cell swelling, while differently regulating cell adhesion. ( A ) Schematic of necrosis and nepoptosis regulation of cell swelling, cell adhesion, and morphological changes measured using a hemocytometer, FACS, the xCELLigence system, and CNT/AFM probes. ( B ) DOX- and ETO-induced cell swelling were visually measured using hemocytometer analysis and detected by phase contrast microscopy (scale bar represents 250 μm) at 72 h. ( C ) DOX- and ETO-induced numerical cell size measured using FSC units of analysis for increasing times. ( D ) DOX- (red color) and ETO (green color)-treated cells were harvested after 72 h and cell adhesion was measured, including cell index (CI) and saturation times (ST) detected using xCELLigence for 10 s intervals and monitored for 3 h. Black closed diagram indicate saturation (±1.0%) times. ( E , F ) Table shows the cell size (FSC), adhesion area (CI), and saturation times (ST; h) at which cell adhesion was examined using computational analysis (described in results) compared to the control. The histogram shows cell adhesion levels calculated using the derived formula (described in results) for DOX- and ETO-treated cells for 72 h. ( G ) DOX-and ETO-treated cells were seeded in culture dishes for 3 h. Expression level of F-actin (red) was measured using immunofluorescence staining and detected by confocal microscopy. Hoechst 33258 (blue) was used for nuclear staining and scale bar represents 50 μm. All histograms represent statistical analysis (P-value of *P

    Journal: Scientific Reports

    Article Title: Structural and functional analysis of cell adhesion and nuclear envelope nano-topography in cell death

    doi: 10.1038/srep15623

    Figure Lengend Snippet: Doxorubicin (DOX) and etoposide (ETO) induced cell swelling, while differently regulating cell adhesion. ( A ) Schematic of necrosis and nepoptosis regulation of cell swelling, cell adhesion, and morphological changes measured using a hemocytometer, FACS, the xCELLigence system, and CNT/AFM probes. ( B ) DOX- and ETO-induced cell swelling were visually measured using hemocytometer analysis and detected by phase contrast microscopy (scale bar represents 250 μm) at 72 h. ( C ) DOX- and ETO-induced numerical cell size measured using FSC units of analysis for increasing times. ( D ) DOX- (red color) and ETO (green color)-treated cells were harvested after 72 h and cell adhesion was measured, including cell index (CI) and saturation times (ST) detected using xCELLigence for 10 s intervals and monitored for 3 h. Black closed diagram indicate saturation (±1.0%) times. ( E , F ) Table shows the cell size (FSC), adhesion area (CI), and saturation times (ST; h) at which cell adhesion was examined using computational analysis (described in results) compared to the control. The histogram shows cell adhesion levels calculated using the derived formula (described in results) for DOX- and ETO-treated cells for 72 h. ( G ) DOX-and ETO-treated cells were seeded in culture dishes for 3 h. Expression level of F-actin (red) was measured using immunofluorescence staining and detected by confocal microscopy. Hoechst 33258 (blue) was used for nuclear staining and scale bar represents 50 μm. All histograms represent statistical analysis (P-value of *P

    Article Snippet: Hoechst 33258 reagent (5 μM; Sigma-Aldrich Co. LLC) for nuclear staining was added to the samples for 30 min at room temperature, and samples were washed with PBS for 3 times.

    Techniques: FACS, Microscopy, Derivative Assay, Expressing, Immunofluorescence, Staining, Confocal Microscopy, Significance Assay

    Drp-1 siRNA prevents Mn damage. Western blot analysis of Drp-1 protein in C6 cells transfected with pooled Drp-1 or control siRNA for 48 hours. Reprobing with an anti-β-Actin antibody was performed to normalize for protein loading. Results are expressed as a percentage of the respective control considered as 100%. Viability was measured by MTT assay ( A ); Quantification of mitochondrial morphology and Δψm dissipation analysis using MitoTracker Red CMXRos (75 nM) ( B ); Apoptotic nuclei were determined by Hoechst 33258 staining (arrowheads). Scale bar: 10 µm ( C ). Results are average of three individual experiments. Statistical significance, **p

    Journal: PLoS ONE

    Article Title: Deregulation of Mitochondria-Shaping Proteins Opa-1 and Drp-1 in Manganese-Induced Apoptosis

    doi: 10.1371/journal.pone.0091848

    Figure Lengend Snippet: Drp-1 siRNA prevents Mn damage. Western blot analysis of Drp-1 protein in C6 cells transfected with pooled Drp-1 or control siRNA for 48 hours. Reprobing with an anti-β-Actin antibody was performed to normalize for protein loading. Results are expressed as a percentage of the respective control considered as 100%. Viability was measured by MTT assay ( A ); Quantification of mitochondrial morphology and Δψm dissipation analysis using MitoTracker Red CMXRos (75 nM) ( B ); Apoptotic nuclei were determined by Hoechst 33258 staining (arrowheads). Scale bar: 10 µm ( C ). Results are average of three individual experiments. Statistical significance, **p

    Article Snippet: Reagents Dulbecco’s Modified Eagle’s Medium (DMEM), manganese chloride, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), Hoechst 33258 fluorochrome, ECL detection reagents (luminol and p-coumaric acid) and 3,3′-diaminobenzidine (DAB) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques: Western Blot, Transfection, MTT Assay, Staining

    WT and Q297V Opa-1 over-expression prevent apoptotic nuclei appearance. Normal and apoptotic (condensed and fragmented chromatin) (arrowheads) nuclei were detected with Hoechst 33258 (1 µg/ml) by fluorescence microscopy and 200 cells/sample were scored. Scale bar: 10 µm. Two independent experiments were conducted. Statistical significance, **p

    Journal: PLoS ONE

    Article Title: Deregulation of Mitochondria-Shaping Proteins Opa-1 and Drp-1 in Manganese-Induced Apoptosis

    doi: 10.1371/journal.pone.0091848

    Figure Lengend Snippet: WT and Q297V Opa-1 over-expression prevent apoptotic nuclei appearance. Normal and apoptotic (condensed and fragmented chromatin) (arrowheads) nuclei were detected with Hoechst 33258 (1 µg/ml) by fluorescence microscopy and 200 cells/sample were scored. Scale bar: 10 µm. Two independent experiments were conducted. Statistical significance, **p

    Article Snippet: Reagents Dulbecco’s Modified Eagle’s Medium (DMEM), manganese chloride, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), Hoechst 33258 fluorochrome, ECL detection reagents (luminol and p-coumaric acid) and 3,3′-diaminobenzidine (DAB) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques: Over Expression, Fluorescence, Microscopy

    Mdivi-1 protects from Mn injury. Viability was measured by MTT assay ( A ); Quantification of mitochondrial morphology and Δψm dissipation analysis using MitoTracker Red CMXRos (75 nM) ( B ); Apoptotic nuclei were determined by staining with Hoechst 33258 (arrowheads). Scale bar: 10 µm ( C ). Results are average of three individual experiments. Statistical significance, **p

    Journal: PLoS ONE

    Article Title: Deregulation of Mitochondria-Shaping Proteins Opa-1 and Drp-1 in Manganese-Induced Apoptosis

    doi: 10.1371/journal.pone.0091848

    Figure Lengend Snippet: Mdivi-1 protects from Mn injury. Viability was measured by MTT assay ( A ); Quantification of mitochondrial morphology and Δψm dissipation analysis using MitoTracker Red CMXRos (75 nM) ( B ); Apoptotic nuclei were determined by staining with Hoechst 33258 (arrowheads). Scale bar: 10 µm ( C ). Results are average of three individual experiments. Statistical significance, **p

    Article Snippet: Reagents Dulbecco’s Modified Eagle’s Medium (DMEM), manganese chloride, 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), Hoechst 33258 fluorochrome, ECL detection reagents (luminol and p-coumaric acid) and 3,3′-diaminobenzidine (DAB) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques: MTT Assay, Staining

    The proliferation of BMSCs in the NF PLLA scaffolds. Rabbit BMSCs were seeded into the porous PLLA scaffolds. After in culture for 1, 3, and 7 days, the cell/scaffold constructs were harvested and used to quantify cell proliferation (DNA content) with the Hoechst 33258 method. Here, * or # designate data that is significantly different from that of 1d or 3d, respectively.

    Journal: Acta biomaterialia

    Article Title: Suppressing Mesenchymal Stem Cell Hypertrophy and Endochondral Ossification in 3D Cartilage Regeneration with Nanofibrous Poly(L-Lactic Acid) Scaffold and Matrilin-3

    doi: 10.1016/j.actbio.2018.06.027

    Figure Lengend Snippet: The proliferation of BMSCs in the NF PLLA scaffolds. Rabbit BMSCs were seeded into the porous PLLA scaffolds. After in culture for 1, 3, and 7 days, the cell/scaffold constructs were harvested and used to quantify cell proliferation (DNA content) with the Hoechst 33258 method. Here, * or # designate data that is significantly different from that of 1d or 3d, respectively.

    Article Snippet: The GAG content was normalized by total DNA that was measured via Hoechst 33258 assay with calf thymus DNA (Sigma-Aldrich) as a standard.

    Techniques: Construct

    Cellular apoptosis in MKN-28 cells was analyzed by Hoechst 33258 staining and flow cytometry. (A) Apoptotic features were identified by observing chromatin condensation and fragmentation after Hoechst 33258 staining, as indicated by white arrows. (B) Detection of apoptosis via annexin V/PI staining (X-axis, annexin V; Y-axis, PI). The early apoptotic cells were defined as the sum of cells in the lower right quadrant of panel B. In A and B: (a) control; (b) 40μM SIN; (c) 100 mg/l 5-FU; and (d) 20 μM SIN + 50 mg/l 5-FU. (C) The apoptotic rate, as assessed via Hoechst 33258 staining. Bars indicate the mean ± SD (n=3). SIN, sinomenine; 5-FU, 5-fluorouracil; PI, propidium iodide.

    Journal: Oncology Letters

    Article Title: Sinomenine sensitizes gastric cancer cells to 5-fluorouracil in vitro and in vivo

    doi: 10.3892/ol.2013.1592

    Figure Lengend Snippet: Cellular apoptosis in MKN-28 cells was analyzed by Hoechst 33258 staining and flow cytometry. (A) Apoptotic features were identified by observing chromatin condensation and fragmentation after Hoechst 33258 staining, as indicated by white arrows. (B) Detection of apoptosis via annexin V/PI staining (X-axis, annexin V; Y-axis, PI). The early apoptotic cells were defined as the sum of cells in the lower right quadrant of panel B. In A and B: (a) control; (b) 40μM SIN; (c) 100 mg/l 5-FU; and (d) 20 μM SIN + 50 mg/l 5-FU. (C) The apoptotic rate, as assessed via Hoechst 33258 staining. Bars indicate the mean ± SD (n=3). SIN, sinomenine; 5-FU, 5-fluorouracil; PI, propidium iodide.

    Article Snippet: Detection of apoptotic cells by Hoechst 33258 staining and flow cytometry The morphological features of apoptotic cells (chromatin condensation and fragmentation) were detected by Hoechst 33258 staining as follows: MKN-28 cells were treated with 100 mg/l 5-FU, 40 μM SIN or 50mg/l 5-FU + 20 μM SIN for 24 h, followed by incubation with 20 μM Hoechst 33258 (Sigma-Aldrich) for 10 min at room temperature.

    Techniques: Staining, Flow Cytometry, Cytometry

    Purified population of oligodendroglial progenitors, used for co‐culture experiments with hippocampal slices. Cell nuclei are visualized by using Hoechst 33258 (blue). (a) Small, round progenitor cells (live), (b) NG2‐positive cells, (c) cells expressing CNP antigen, (d) co‐expression of CNP (red) and PDGFRα (green) antigens. (e) Proliferating (Ki67 + ) cells, (f) Co‐expression of NG2 (green) and NF200 (red) markers, rarely (

    Journal: Cell Proliferation

    Article Title: Crucial role of the local micro‐environment in fate decision of neonatal rat NG2 progenitors

    doi: 10.1111/j.1365-2184.2009.00618.x

    Figure Lengend Snippet: Purified population of oligodendroglial progenitors, used for co‐culture experiments with hippocampal slices. Cell nuclei are visualized by using Hoechst 33258 (blue). (a) Small, round progenitor cells (live), (b) NG2‐positive cells, (c) cells expressing CNP antigen, (d) co‐expression of CNP (red) and PDGFRα (green) antigens. (e) Proliferating (Ki67 + ) cells, (f) Co‐expression of NG2 (green) and NF200 (red) markers, rarely (

    Article Snippet: Cell nuclei were visualized by incubation with 5 µ m Hoechst 33258 (Sigma).

    Techniques: Purification, Co-Culture Assay, Expressing

    Nuclear morphology study for apoptosis. Chondrocytes were incubated for 16 hours with or without 30 μM 4-hydroxynonenal (HNE), stained with Hoechst 33258, and then analyzed by fluorescence microscopy.

    Journal: Arthritis Research & Therapy

    Article Title: 4-Hydroxynonenal induces apoptosis in human osteoarthritic chondrocytes: the protective role of glutathione-S-transferase

    doi: 10.1186/ar2503

    Figure Lengend Snippet: Nuclear morphology study for apoptosis. Chondrocytes were incubated for 16 hours with or without 30 μM 4-hydroxynonenal (HNE), stained with Hoechst 33258, and then analyzed by fluorescence microscopy.

    Article Snippet: Nuclear morphology study for apoptosis Apoptotic nuclear morphology was assessed by Hoechst 33258 incorporation (Sigma-Aldrich).

    Techniques: Incubation, Staining, Fluorescence, Microscopy

    Geminin interacts with TopoIIα on chromosomes in HME cells . (A) HME cells synchronized in G 0 /G 1 , S, G 2 /M and M/G 1 were sonicated and immunoprecipitated with the indicated antibodies, followed by immunoblotting with the indicated antibodies. (B) HME cells treated with etoposide (10 μM, overnight) were then collected and processed for analysis using trapped in agarose DNA immunostaining (TARDIS) assay. Shown are examples of cells costained with Hoechst 33258 blue, TopoIIα and geminin antibodies (1 to 3), costained with Hoechst 33258 blue, TopoIIα but not Cdc7 antibodies (4 to 6) or costained with Hoechst 33258 blue, geminin but not Cdc7 antibodies (7 to 9). Geminin-silenced HME cells (10), plus cell division cycle 7 silencing (siCdc7) (11) or plus casein kinase Iε (CKIε) overexpression (12) are shown. TopoIIα-silenced HME cells (13) plus Cdc7-silenced (14) or plus CKIε overexpression (15) are also shown. (C) Quantification of cells costained with Hoechst 33258 blue and Cdc7 (white bars), geminin (red bars) or TopoIIα (black bars) in TARDIS assays 72 hours after control, Cdc7, geminin, TopoIIα, geminin silencing and Cdc7 silencing (or CKIε overexpression); geminin and TopoIIα silencing; or TopoIIα silencing and Cdc7 silencing (or CKIε overexpression). Values presented are means ± SD. ** P ≤ 0.01. (D) In vitro kinase assay performed on purified TopoIIα using CKIε or Cdc7 immunoprecipitated from HME cells.

    Journal: Breast Cancer Research : BCR

    Article Title: Geminin overexpression prevents the completion of topoisomerase II? chromosome decatenation, leading to aneuploidy in human mammary epithelial cells

    doi: 10.1186/bcr2884

    Figure Lengend Snippet: Geminin interacts with TopoIIα on chromosomes in HME cells . (A) HME cells synchronized in G 0 /G 1 , S, G 2 /M and M/G 1 were sonicated and immunoprecipitated with the indicated antibodies, followed by immunoblotting with the indicated antibodies. (B) HME cells treated with etoposide (10 μM, overnight) were then collected and processed for analysis using trapped in agarose DNA immunostaining (TARDIS) assay. Shown are examples of cells costained with Hoechst 33258 blue, TopoIIα and geminin antibodies (1 to 3), costained with Hoechst 33258 blue, TopoIIα but not Cdc7 antibodies (4 to 6) or costained with Hoechst 33258 blue, geminin but not Cdc7 antibodies (7 to 9). Geminin-silenced HME cells (10), plus cell division cycle 7 silencing (siCdc7) (11) or plus casein kinase Iε (CKIε) overexpression (12) are shown. TopoIIα-silenced HME cells (13) plus Cdc7-silenced (14) or plus CKIε overexpression (15) are also shown. (C) Quantification of cells costained with Hoechst 33258 blue and Cdc7 (white bars), geminin (red bars) or TopoIIα (black bars) in TARDIS assays 72 hours after control, Cdc7, geminin, TopoIIα, geminin silencing and Cdc7 silencing (or CKIε overexpression); geminin and TopoIIα silencing; or TopoIIα silencing and Cdc7 silencing (or CKIε overexpression). Values presented are means ± SD. ** P ≤ 0.01. (D) In vitro kinase assay performed on purified TopoIIα using CKIε or Cdc7 immunoprecipitated from HME cells.

    Article Snippet: Slides were counterstained with Hoechst 33258 blue (10 μM in PBS; Sigma) for 5 minutes before application of coverslips that were secured with a sealant.

    Techniques: Sonication, Immunoprecipitation, Immunostaining, Over Expression, In Vitro, Kinase Assay, Purification

    Hoechst 33258 staining indicated that GAS inhibits H 2 O 2 -induced HepG2 cells apoptosis. Morphologic changes in nuclei observed with Hoechst 33258 staining under fluorescence microscopy. (a) Natural group. The HepG2 cells showed normal shape with round intact nuclei; (b) H 2 O 2 group, HepG2 cells treated with 1600 μ M H 2 O 2 for 4 hours, and the obvious morphologic changes were observed; (c) H 2 O 2 + Vitamin C (200 nM) group; (d) H 2 O 2 + GAS (125 μ g/mL) group; (e) H 2 O 2 + GAS (25 μ g/mL) group; (f) H 2 O 2 + GAS (5 μ g/mL) group. The arrows indicate apoptotic cells. Original magnification is 400x.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: The Antiapoptosis Effect of Glycyrrhizate on HepG2 Cells Induced by Hydrogen Peroxide

    doi: 10.1155/2016/6849758

    Figure Lengend Snippet: Hoechst 33258 staining indicated that GAS inhibits H 2 O 2 -induced HepG2 cells apoptosis. Morphologic changes in nuclei observed with Hoechst 33258 staining under fluorescence microscopy. (a) Natural group. The HepG2 cells showed normal shape with round intact nuclei; (b) H 2 O 2 group, HepG2 cells treated with 1600 μ M H 2 O 2 for 4 hours, and the obvious morphologic changes were observed; (c) H 2 O 2 + Vitamin C (200 nM) group; (d) H 2 O 2 + GAS (125 μ g/mL) group; (e) H 2 O 2 + GAS (25 μ g/mL) group; (f) H 2 O 2 + GAS (5 μ g/mL) group. The arrows indicate apoptotic cells. Original magnification is 400x.

    Article Snippet: Hoechst 33258 stains were purchased from Sigma-Aldrich, USA.

    Techniques: Staining, Fluorescence, Microscopy

    Coxiella OmpA is involved in host cell invasion and intracellular replication. A431 ( A ) and THP-1 ( E ) cells were incubated with wt Coxiella , the control transposon mutant Tn1832 or the OmpA mutant Tn208 . 60 minutes after infection (top panels) cells were fixed and labeled with an anti- Coxiella antibody coupled to Alexa Fluor 555 (blue) and with Atto-647N phalloidin (red) prior to cell permeabilization. Internalized bacteria were detected by GFP fluorescence (green) in the case of Tn208 and Tn1832 whereas for wt Coxiella infections, cells were permeabilized and bacteria were stained with the anti- Coxiella antibody as above, coupled to Alexa Fluor 488 (green). Alternatively, cells were fixed 6 ( A , bottom panels) or 5 ( E , bottom panels) days after infection, DNA (red) was labeled with Hoechst 33258 and wt Coxiella (green) with the specific antibody as above. The automated image analysis software CellProfiler was used to calculate the percentage of internalized bacteria ( B and F ), the number of colonies/cell ( C and G ) and the area (in microns 2 ) of intracellular Coxiella colonies identified for each condition ( D and H ). Values are means ± standard deviations of triplicate experiments where an average of 8000 bacteria (B and F) or 400 vacuoles (C, D, G, H) were analyzed for each condition (values were compared to wt Coxiella infections. ns = non-significant; *** = P

    Journal: PLoS Pathogens

    Article Title: Identification of OmpA, a Coxiella burnetii Protein Involved in Host Cell Invasion, by Multi-Phenotypic High-Content Screening

    doi: 10.1371/journal.ppat.1004013

    Figure Lengend Snippet: Coxiella OmpA is involved in host cell invasion and intracellular replication. A431 ( A ) and THP-1 ( E ) cells were incubated with wt Coxiella , the control transposon mutant Tn1832 or the OmpA mutant Tn208 . 60 minutes after infection (top panels) cells were fixed and labeled with an anti- Coxiella antibody coupled to Alexa Fluor 555 (blue) and with Atto-647N phalloidin (red) prior to cell permeabilization. Internalized bacteria were detected by GFP fluorescence (green) in the case of Tn208 and Tn1832 whereas for wt Coxiella infections, cells were permeabilized and bacteria were stained with the anti- Coxiella antibody as above, coupled to Alexa Fluor 488 (green). Alternatively, cells were fixed 6 ( A , bottom panels) or 5 ( E , bottom panels) days after infection, DNA (red) was labeled with Hoechst 33258 and wt Coxiella (green) with the specific antibody as above. The automated image analysis software CellProfiler was used to calculate the percentage of internalized bacteria ( B and F ), the number of colonies/cell ( C and G ) and the area (in microns 2 ) of intracellular Coxiella colonies identified for each condition ( D and H ). Values are means ± standard deviations of triplicate experiments where an average of 8000 bacteria (B and F) or 400 vacuoles (C, D, G, H) were analyzed for each condition (values were compared to wt Coxiella infections. ns = non-significant; *** = P

    Article Snippet: Antibodies and reagents Hoechst 33258, rabbit anti poly-His, anti mouse and anti-rabbit HRP-conjugated antibodies and Atto-647N phalloidin were purchased from Sigma.

    Techniques: Incubation, Mutagenesis, Infection, Labeling, Fluorescence, Staining, Software

    Expression of MMP-2 and comparisons of apoptotic indexes among different groups. Top panel, there was much stronger MMP-2 staining in tumor tissue of VEGFD-SK group compared with the weak staining in WT-SK group and EV-SK group. The pVAX:lip group also exhibited deep MMP-2 staining; whereas the pVAX-MP:lip group showed negative MMP-2 staining. Middle panel, microscopic fluorescence images show apoptotic cells of tumor tissues of different groups by Hoechst-33258 staining. VEGFD-SK tumor tissue manifested a few apoptosis of tumor cells; whereas pVAX-MP:lip group tumor tissue revealed a significant enhancement in apoptosis of tumor cells (arrow, apoptotic cell). However, no appreciable difference was observed between WT-SK group and EV-SK group. Bottom panel, VEGFD-SK group tumor tissue manifested a significant suppression of apoptotic tumor cells in contrast to other groups. Whereas higher apoptosis index was observed after the pVAX-MP:lip administration ( * P

    Journal: International Journal of Oncology

    Article Title: VEGF-D-enhanced lymph node metastasis of ovarian cancer is reversed by vesicular stomatitis virus matrix protein

    doi: 10.3892/ijo.2016.3527

    Figure Lengend Snippet: Expression of MMP-2 and comparisons of apoptotic indexes among different groups. Top panel, there was much stronger MMP-2 staining in tumor tissue of VEGFD-SK group compared with the weak staining in WT-SK group and EV-SK group. The pVAX:lip group also exhibited deep MMP-2 staining; whereas the pVAX-MP:lip group showed negative MMP-2 staining. Middle panel, microscopic fluorescence images show apoptotic cells of tumor tissues of different groups by Hoechst-33258 staining. VEGFD-SK tumor tissue manifested a few apoptosis of tumor cells; whereas pVAX-MP:lip group tumor tissue revealed a significant enhancement in apoptosis of tumor cells (arrow, apoptotic cell). However, no appreciable difference was observed between WT-SK group and EV-SK group. Bottom panel, VEGFD-SK group tumor tissue manifested a significant suppression of apoptotic tumor cells in contrast to other groups. Whereas higher apoptosis index was observed after the pVAX-MP:lip administration ( * P

    Article Snippet: Apoptosis analysis Qualitative apoptosis was determined by Hoechst-33258 compounds (Sigma) according to the manufacturer's instructions.

    Techniques: Expressing, Staining, Fluorescence

    RT-PCR and apoptotic effect on VEGFD-SK cells. (A) RT-PCR analysis of expression of MP in vitro . High expression of MP in VEGFD-SK cells transfected with pVAX-MP:lip was detected (1,036 bp). GAPDH was used for the internal standard. The displaying panel of electrophoretic image presents the following: DNA ladder marker (a), WT-SK (b), EV-SK (c), VEGFD-SK (d), pVAX:lip (e), and pVAX-MP:lip (f), respectively. (B) Overexpression of VEGF-D using RT-PCR could be detected in VEGFD-SK cells, but not in WT-SK and EV-SK cells (1,077 bp). However, the VEGF-D expression was suppressed when VEGFD-SK cells suffered from pVAX-MP:lip. The overexpression of VEGF-D did not decreased obviously when VEGFD-SK cells were treated with pVA. (C) VEGF-D expression in vivo was analysis by RT-PCR. (D) The expression of VEGF-D in vivo was determined with immunohistochemistry using VEGF-D antibody: VEGFD-SK tumors were detected with much stronger staining. Whereas, the VEGFD-SK tumors in pVAX-MP:lip group present nearly negative staining. Tumors in pVAX:lip group also exhibited strong staining. Very weak VEGF-D protein was observed in WT-SK and EV-SK tumors. (E) Hoechst-33258 stained fluorescence microscopy (x200) in vitro : apoptosis cells were observed apparently when the VEGFD-SK cells were treated with pVAX-MP:lip (arrow). (F) A quantitative comparison of apoptotic cells was carried out by flow analysis stained with Annexin V-FITC and propidium iodide: pVAX-MP significantly increased VEGFD-SK cells apoptosis compared with the controls.

    Journal: International Journal of Oncology

    Article Title: VEGF-D-enhanced lymph node metastasis of ovarian cancer is reversed by vesicular stomatitis virus matrix protein

    doi: 10.3892/ijo.2016.3527

    Figure Lengend Snippet: RT-PCR and apoptotic effect on VEGFD-SK cells. (A) RT-PCR analysis of expression of MP in vitro . High expression of MP in VEGFD-SK cells transfected with pVAX-MP:lip was detected (1,036 bp). GAPDH was used for the internal standard. The displaying panel of electrophoretic image presents the following: DNA ladder marker (a), WT-SK (b), EV-SK (c), VEGFD-SK (d), pVAX:lip (e), and pVAX-MP:lip (f), respectively. (B) Overexpression of VEGF-D using RT-PCR could be detected in VEGFD-SK cells, but not in WT-SK and EV-SK cells (1,077 bp). However, the VEGF-D expression was suppressed when VEGFD-SK cells suffered from pVAX-MP:lip. The overexpression of VEGF-D did not decreased obviously when VEGFD-SK cells were treated with pVA. (C) VEGF-D expression in vivo was analysis by RT-PCR. (D) The expression of VEGF-D in vivo was determined with immunohistochemistry using VEGF-D antibody: VEGFD-SK tumors were detected with much stronger staining. Whereas, the VEGFD-SK tumors in pVAX-MP:lip group present nearly negative staining. Tumors in pVAX:lip group also exhibited strong staining. Very weak VEGF-D protein was observed in WT-SK and EV-SK tumors. (E) Hoechst-33258 stained fluorescence microscopy (x200) in vitro : apoptosis cells were observed apparently when the VEGFD-SK cells were treated with pVAX-MP:lip (arrow). (F) A quantitative comparison of apoptotic cells was carried out by flow analysis stained with Annexin V-FITC and propidium iodide: pVAX-MP significantly increased VEGFD-SK cells apoptosis compared with the controls.

    Article Snippet: Apoptosis analysis Qualitative apoptosis was determined by Hoechst-33258 compounds (Sigma) according to the manufacturer's instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, In Vitro, Transfection, Marker, Over Expression, In Vivo, Immunohistochemistry, Staining, Negative Staining, Fluorescence, Microscopy, Flow Cytometry

    Proliferation of rASCs cultured under different conditions determined by DNA assay using Hoechst 33258 dye. rASCs group was cultured with LG-DMEM+10% FBS; BM group: LG-DMEM+2% FBS; RHE-treated group: LG-DMEM+2% FBS+2.5 μM ATRA+20 ng/mL

    Journal: Tissue Engineering. Part A

    Article Title: Effects of Multiple Agents on Epithelial Differentiation of Rabbit Adipose-Derived Stem Cells in 3D Culture

    doi: 10.1089/ten.tea.2011.0424

    Figure Lengend Snippet: Proliferation of rASCs cultured under different conditions determined by DNA assay using Hoechst 33258 dye. rASCs group was cultured with LG-DMEM+10% FBS; BM group: LG-DMEM+2% FBS; RHE-treated group: LG-DMEM+2% FBS+2.5 μM ATRA+20 ng/mL

    Article Snippet: The resulting mixture was subjected to centrifugation and aliquots of the supernatants after mixing with 160 μL Hoechst 33258 dye solution (0.1 μg/mL; Sigma-Aldrich) were transferred to black flat-bottomed 96-well plates (Corning Costar).

    Techniques: Cell Culture

    Effect of RNA interference (RNAi) on apoptosis and survival rate of Parkinson's disease model cells. (A–C) Morphological changes to apoptotic cells that appeared in model and RNAi groups were detected by Hoechst 33258 staining (fluorescence inverted microscope, ×400); apoptotic cells were stained as blue. (D) Apoptosis rate was detected by flow cytometry. (E) Cell survival rate was de-tected by MTT assay. The rate of apoptosis and survival rate in Parkinson's disease cells in the model group was not significantly different from that in the RNAi group. Data are expressed as mean ± SD and were analyzed using analysis of variance. a P

    Journal: Neural Regeneration Research

    Article Title: The Pael-R gene does not mediate the changes in rotenone-induced Parkinson's disease model cells

    doi: 10.4103/1673-5374.128245

    Figure Lengend Snippet: Effect of RNA interference (RNAi) on apoptosis and survival rate of Parkinson's disease model cells. (A–C) Morphological changes to apoptotic cells that appeared in model and RNAi groups were detected by Hoechst 33258 staining (fluorescence inverted microscope, ×400); apoptotic cells were stained as blue. (D) Apoptosis rate was detected by flow cytometry. (E) Cell survival rate was de-tected by MTT assay. The rate of apoptosis and survival rate in Parkinson's disease cells in the model group was not significantly different from that in the RNAi group. Data are expressed as mean ± SD and were analyzed using analysis of variance. a P

    Article Snippet: Cell morphological changes tested by Hoechst 33258 staining Hoechst 33258 solution (Sigma) was added into the three groups of cultured cells to a final concentration of 10 μg/mL; then, cells were incubated for 5 minutes at room temperature.

    Techniques: Staining, Fluorescence, Inverted Microscopy, Flow Cytometry, Cytometry, MTT Assay

    PNP-AV membrane binding visualization. Confocal microscopy of MCF-7 cells confirms the presence of externally bound biotinylated PNP-AV, with streptavidin-conjugated Alexa Fluor 488 in green (a, e), red CellMask stain of the plasma membrane (b, f), and blue Hoechst 33258 dye staining of nucleic acids (c, g). Images a-c show the color channels of an x-y confocal image with the composite shown in d. Images e-g show the color channels of an x-z cross-section of cells with the composite shown in h and the coverslip indicated by the white in each image.

    Journal: PLoS ONE

    Article Title: Purine Nucleoside Phosphorylase Targeted by Annexin V to Breast Cancer Vasculature for Enzyme Prodrug Therapy

    doi: 10.1371/journal.pone.0076403

    Figure Lengend Snippet: PNP-AV membrane binding visualization. Confocal microscopy of MCF-7 cells confirms the presence of externally bound biotinylated PNP-AV, with streptavidin-conjugated Alexa Fluor 488 in green (a, e), red CellMask stain of the plasma membrane (b, f), and blue Hoechst 33258 dye staining of nucleic acids (c, g). Images a-c show the color channels of an x-y confocal image with the composite shown in d. Images e-g show the color channels of an x-z cross-section of cells with the composite shown in h and the coverslip indicated by the white in each image.

    Article Snippet: Cells were counterstained with 5 µg/ml Hoechst 33258 blue dye (Sigma-Aldrich) for nucleic acids for 30 min and with 1 µg/ml CellMask Deep Red plasma membrane stain (Invitrogen) using three 5 min incubations followed by washes.

    Techniques: Binding Assay, Confocal Microscopy, Staining

    Immunofluorescence labeling with three different mAbs directed against rat postnatal cortical membrane preparations in the developing rat cortex. A , mab 10 ; B , mab 11 ; C , mab 942 . Top , Frontal sections from P6 cortex; bottom , frontal sections from E19 cortex. To the left of each fluorescent image is a micrograph of the left half of the same section counterstained with bisbenzimide to illustrate the cortical layering. MZ , Marginal zone; CP , cortical plate; SP , subplate zone; IZ ; intermediate zone; VZ , ventricular zone. Scale bars, 100 μm.

    Journal: The Journal of Neuroscience

    Article Title: Dual Action of a Carbohydrate Epitope on Afferent and Efferent Axons in Cortical Development

    doi: 10.1523/JNEUROSCI.16-13-04195.1996

    Figure Lengend Snippet: Immunofluorescence labeling with three different mAbs directed against rat postnatal cortical membrane preparations in the developing rat cortex. A , mab 10 ; B , mab 11 ; C , mab 942 . Top , Frontal sections from P6 cortex; bottom , frontal sections from E19 cortex. To the left of each fluorescent image is a micrograph of the left half of the same section counterstained with bisbenzimide to illustrate the cortical layering. MZ , Marginal zone; CP , cortical plate; SP , subplate zone; IZ ; intermediate zone; VZ , ventricular zone. Scale bars, 100 μm.

    Article Snippet: To label cell nuclei, sections were counterstained with bisbenzimide solution (1 μg/ml in PBS, 5 min at room temperature; Sigma, St. Louis, MO).

    Techniques: Immunofluorescence, Labeling