Structured Review

Becton Dickinson facscan flow cytometer
SC-58125 does not induce apoptosis. LLC (A) or HCA-7 (B) cells untreated (negative control), treated with SC-58635 (positive control), DMSO (vehicle), or with 100 µM SC-58125 for 24 hours (LLC), or 72 hours (HCA-7). Cells were harvested then fixed, and fragmented DNA was labeled using the TUNEL assay with dUTP- FITC as substrate. Analysis was performed using a Becton Dickinson <t>FACScan</t> flow cytometer. Ten thousand gated events were analyzed.
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Images

1) Product Images from "A Cyclooxygenase-2 Inhibitor (SC-58125) Blocks Growth of Established Human Colon Cancer Xenografts 1"

Article Title: A Cyclooxygenase-2 Inhibitor (SC-58125) Blocks Growth of Established Human Colon Cancer Xenografts 1

Journal: Neoplasia (New York, N.Y.)

doi:

SC-58125 does not induce apoptosis. LLC (A) or HCA-7 (B) cells untreated (negative control), treated with SC-58635 (positive control), DMSO (vehicle), or with 100 µM SC-58125 for 24 hours (LLC), or 72 hours (HCA-7). Cells were harvested then fixed, and fragmented DNA was labeled using the TUNEL assay with dUTP- FITC as substrate. Analysis was performed using a Becton Dickinson FACScan flow cytometer. Ten thousand gated events were analyzed.
Figure Legend Snippet: SC-58125 does not induce apoptosis. LLC (A) or HCA-7 (B) cells untreated (negative control), treated with SC-58635 (positive control), DMSO (vehicle), or with 100 µM SC-58125 for 24 hours (LLC), or 72 hours (HCA-7). Cells were harvested then fixed, and fragmented DNA was labeled using the TUNEL assay with dUTP- FITC as substrate. Analysis was performed using a Becton Dickinson FACScan flow cytometer. Ten thousand gated events were analyzed.

Techniques Used: High Content Screening, Negative Control, Positive Control, Labeling, TUNEL Assay, Flow Cytometry, Cytometry

2) Product Images from "A Precise Nanostructure of Folate-Overhung Mitoxantrone DNA Tetrahedron for Targeted Capture Leukemia"

Article Title: A Precise Nanostructure of Folate-Overhung Mitoxantrone DNA Tetrahedron for Targeted Capture Leukemia

Journal: Nanomaterials

doi: 10.3390/nano10050951

Induced apoptosis in leukemia BALL-1 cells by folate-overhung mitoxantrone DNA tetrahedra. ( A–D ): Induced apoptosis after treatment with varying formulations at 10 h, and the cells were stained using Annexin V-Fluor 647 and 7AAD, then assessed according to manufacturer instructions by the FACScan flow cytometer. UR: Late apoptosis and dead cells; LR: Early apoptosis cells ( A ). PBS; ( B ). Free mitoxantrone; ( C ). mitoxantrone DNA tetrahedra; ( D ). Folate-overhung mitoxantrone DNA tetrahedra; ( E ). The early apoptosis rate in BALL-1 cells ( F ). The late apoptosis rate in BALL-1 cells. ( E–F ): 1. PBS; 2. Free mitoxantrone; 3. Mitoxantrone DNA tetrahedra; 4. Folate-overhung mitoxantrone DNA tetrahedra. a. vs. PBS; b. vs. mitoxantrone DNA tetrahedra, p
Figure Legend Snippet: Induced apoptosis in leukemia BALL-1 cells by folate-overhung mitoxantrone DNA tetrahedra. ( A–D ): Induced apoptosis after treatment with varying formulations at 10 h, and the cells were stained using Annexin V-Fluor 647 and 7AAD, then assessed according to manufacturer instructions by the FACScan flow cytometer. UR: Late apoptosis and dead cells; LR: Early apoptosis cells ( A ). PBS; ( B ). Free mitoxantrone; ( C ). mitoxantrone DNA tetrahedra; ( D ). Folate-overhung mitoxantrone DNA tetrahedra; ( E ). The early apoptosis rate in BALL-1 cells ( F ). The late apoptosis rate in BALL-1 cells. ( E–F ): 1. PBS; 2. Free mitoxantrone; 3. Mitoxantrone DNA tetrahedra; 4. Folate-overhung mitoxantrone DNA tetrahedra. a. vs. PBS; b. vs. mitoxantrone DNA tetrahedra, p

Techniques Used: Staining, Flow Cytometry

3) Product Images from "Mechanisms Involved in Stimulation of Human Immunodeficiency Virus Type 1 Replication by Aminooxypentane RANTES"

Article Title: Mechanisms Involved in Stimulation of Human Immunodeficiency Virus Type 1 Replication by Aminooxypentane RANTES

Journal: Journal of Virology

doi: 10.1128/JVI.75.18.8624-8638.2001

Surface expression of CCR5 and CXCR4 in PBMC treated with AOP-RANTES and RANTES. PBMC were exposed to AOP-RANTES or RANTES for 12 h and then costained with PerCP-labeled anti-CD4 antibody and PE-labeled anti-CCR5 or anti-CXCR4 antibody. Untreated PBMC were also labeled with the anti-CD4 antibody and a conjugated mouse IgG2a κ isotype standard. Labeled cells were then analyzed using the FacScan flow cytometer and Lysis II software. Panels I to VI represent CCR5 (A) or CXCR4 (B) events in the CD4 positive-gated lymphocyte population. (C and D) Comparison of CCR5 or CXCR4 surface expression to NSI/R5 B-92BR021 and SI/X4 D-92UG021 virus production (day 10), respectively. P.Tx., pertussis toxin.
Figure Legend Snippet: Surface expression of CCR5 and CXCR4 in PBMC treated with AOP-RANTES and RANTES. PBMC were exposed to AOP-RANTES or RANTES for 12 h and then costained with PerCP-labeled anti-CD4 antibody and PE-labeled anti-CCR5 or anti-CXCR4 antibody. Untreated PBMC were also labeled with the anti-CD4 antibody and a conjugated mouse IgG2a κ isotype standard. Labeled cells were then analyzed using the FacScan flow cytometer and Lysis II software. Panels I to VI represent CCR5 (A) or CXCR4 (B) events in the CD4 positive-gated lymphocyte population. (C and D) Comparison of CCR5 or CXCR4 surface expression to NSI/R5 B-92BR021 and SI/X4 D-92UG021 virus production (day 10), respectively. P.Tx., pertussis toxin.

Techniques Used: Expressing, Labeling, Flow Cytometry, Cytometry, Lysis, Software

4) Product Images from "High-Throughput Multiplex Flow Cytometry Screening for Botulinum Neurotoxin Type A Light Chain Protease Inhibitors"

Article Title: High-Throughput Multiplex Flow Cytometry Screening for Botulinum Neurotoxin Type A Light Chain Protease Inhibitors

Journal: Assay and Drug Development Technologies

doi: 10.1089/adt.2009.0219

Five-plex flow cytometry assay with botulinum neurotoxin type A light chain (BoNTALC) run continuously on a FACScan flow cytometer. ( A ) Multiplex bead set of FL2 vs. number of events showing the 5 distinct populations of microspheres. ( B ) Continuously
Figure Legend Snippet: Five-plex flow cytometry assay with botulinum neurotoxin type A light chain (BoNTALC) run continuously on a FACScan flow cytometer. ( A ) Multiplex bead set of FL2 vs. number of events showing the 5 distinct populations of microspheres. ( B ) Continuously

Techniques Used: Flow Cytometry, Cytometry, Multiplex Assay

5) Product Images from "Apoptosis and apoptosis-associated perturbations of peripheral blood lymphocytes during HIV infection: comparison between AIDS patients and asymptomatic long-term non-progressors"

Article Title: Apoptosis and apoptosis-associated perturbations of peripheral blood lymphocytes during HIV infection: comparison between AIDS patients and asymptomatic long-term non-progressors

Journal: Clinical and Experimental Immunology

doi: 10.1046/j.1365-2249.2000.01375.x

Assessment of superoxide anion generation in peripheral blood T lymphocytes from AIDS patients and long-term non-progressors (LTNPs). Two-colour staining was performed for the simultaneous determination of CD4/CD8 surface phenotype and superoxide anion generation. Cells from representative AIDS patients and LTNPs were labelled, as described in Materials and methods, using markers for superoxide anion generation (hydroxyethidine (HE)) and MoAbs to CD4/CD8 antigens (a/b, respectively). Numbers refer to the percentage of cells bearing an HE→Eth high phenotype among the CD4 + and CD8 + T lymphocytes. In the FACScan profiles, apoptotic cells were identified either by forward light scatter (FSC)/side scatter (SSC) criteria or by positive staining with 7-amino-actinomycin D (7-AAD), as described in Materials and methods. The cursors were set according to the background staining defined with mouse IgG isotypic controls. Results are representative of two independent experiments each performed on two different subjects.
Figure Legend Snippet: Assessment of superoxide anion generation in peripheral blood T lymphocytes from AIDS patients and long-term non-progressors (LTNPs). Two-colour staining was performed for the simultaneous determination of CD4/CD8 surface phenotype and superoxide anion generation. Cells from representative AIDS patients and LTNPs were labelled, as described in Materials and methods, using markers for superoxide anion generation (hydroxyethidine (HE)) and MoAbs to CD4/CD8 antigens (a/b, respectively). Numbers refer to the percentage of cells bearing an HE→Eth high phenotype among the CD4 + and CD8 + T lymphocytes. In the FACScan profiles, apoptotic cells were identified either by forward light scatter (FSC)/side scatter (SSC) criteria or by positive staining with 7-amino-actinomycin D (7-AAD), as described in Materials and methods. The cursors were set according to the background staining defined with mouse IgG isotypic controls. Results are representative of two independent experiments each performed on two different subjects.

Techniques Used: Staining

6) Product Images from "Antimonial-Mediated DNA Fragmentation in Leishmania infantum Amastigotes"

Article Title: Antimonial-Mediated DNA Fragmentation in Leishmania infantum Amastigotes

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.45.7.2064-2069.2001

Cytofluorometry (TUNEL) analysis of the antimonial-induced DNA fragmentation in axenic amastigotes. Results are shown for untreated wild-type parasite control (A) and wild-type amastigotes incubated for 24 h with medium in the presence of 2 mg of geneticin per ml (B), 2 mg of geneticin per ml without terminal transferase enzyme (negative control) (C), 10 and 50 μg of potassium antimonyl tartrate [Sb(III)] per ml (D and E, respectively), or 160 μg of meglumine [Sb(V)] per ml (F). Results for untreated LdiR120 amastigotes (G) and LdiR120 amastigotes incubated in medium containing 50 μg of potassium antimonyl tartrate per ml (H) are shown. After the incubation period, parasites were processed by the TUNEL technique and analyzed on a FACScan (Becton Dickinson). The increase in FL1 fluorescence intensity was monitored. The percentage of apoptotic cells which corresponds to an increase of FL1 fluorescence (M2) is indicated for each experimental condition.
Figure Legend Snippet: Cytofluorometry (TUNEL) analysis of the antimonial-induced DNA fragmentation in axenic amastigotes. Results are shown for untreated wild-type parasite control (A) and wild-type amastigotes incubated for 24 h with medium in the presence of 2 mg of geneticin per ml (B), 2 mg of geneticin per ml without terminal transferase enzyme (negative control) (C), 10 and 50 μg of potassium antimonyl tartrate [Sb(III)] per ml (D and E, respectively), or 160 μg of meglumine [Sb(V)] per ml (F). Results for untreated LdiR120 amastigotes (G) and LdiR120 amastigotes incubated in medium containing 50 μg of potassium antimonyl tartrate per ml (H) are shown. After the incubation period, parasites were processed by the TUNEL technique and analyzed on a FACScan (Becton Dickinson). The increase in FL1 fluorescence intensity was monitored. The percentage of apoptotic cells which corresponds to an increase of FL1 fluorescence (M2) is indicated for each experimental condition.

Techniques Used: TUNEL Assay, Incubation, Negative Control, Fluorescence

7) Product Images from "Sustained Elevated Levels of VCAM-1 in Cultured Fibroblast-like Synoviocytes Can Be Achieved by TNF-? in Combination with Either IL-4 or IL-13 through Increased mRNA Stability"

Article Title: Sustained Elevated Levels of VCAM-1 in Cultured Fibroblast-like Synoviocytes Can Be Achieved by TNF-? in Combination with Either IL-4 or IL-13 through Increased mRNA Stability

Journal: The American Journal of Pathology

doi:

Elevated expression of VCAM-1 cell surface protein ( A ) and mRNA ( B ) by freshly isolated rheumatoid fibroblast-like synoviocytes declines with increasing time in culture. FLSs were isolated as described in Methods and cultured with 10% FBS for various periods of time. Medium was replenished every 5 days. A: cell surface VCAM-1 expression was analyzed after 1, 2, 4, and 6 weeks by flow cytometry using mAb BBA-5. Samples were analyzed using a FACScan flow cytometer and the results expressed as corrected mean fluorescence intensity. Data are presented as the mean ± SE of four experiments. * indicates p
Figure Legend Snippet: Elevated expression of VCAM-1 cell surface protein ( A ) and mRNA ( B ) by freshly isolated rheumatoid fibroblast-like synoviocytes declines with increasing time in culture. FLSs were isolated as described in Methods and cultured with 10% FBS for various periods of time. Medium was replenished every 5 days. A: cell surface VCAM-1 expression was analyzed after 1, 2, 4, and 6 weeks by flow cytometry using mAb BBA-5. Samples were analyzed using a FACScan flow cytometer and the results expressed as corrected mean fluorescence intensity. Data are presented as the mean ± SE of four experiments. * indicates p

Techniques Used: Expressing, Isolation, Cell Culture, Flow Cytometry, Cytometry, Fluorescence

Flow-cytometric analysis of VCAM-1 on RA fibroblast-like synoviocytes treated with TNF-α and IL-4. Cells were cultured with either medium only (untreated), TNF-α (10 ng/ml), IL-4 (10 ng/ml), or TNF-α plus IL-4 for 3 days. Flow cytometry for cell surface VCAM-1 was carried out using mAb BBA-5 as primary antibody. Samples were analyzed using a FACScan flow cytometer. The dashed histogram (in untreated cells) represents staining with PE-conjugated anti-mouse IgG only. Solid line histograms represent cells stained with anti-VCAM-1 and PE-conjugated anti-mouse IgG. Results shown are representative of four experiments.
Figure Legend Snippet: Flow-cytometric analysis of VCAM-1 on RA fibroblast-like synoviocytes treated with TNF-α and IL-4. Cells were cultured with either medium only (untreated), TNF-α (10 ng/ml), IL-4 (10 ng/ml), or TNF-α plus IL-4 for 3 days. Flow cytometry for cell surface VCAM-1 was carried out using mAb BBA-5 as primary antibody. Samples were analyzed using a FACScan flow cytometer. The dashed histogram (in untreated cells) represents staining with PE-conjugated anti-mouse IgG only. Solid line histograms represent cells stained with anti-VCAM-1 and PE-conjugated anti-mouse IgG. Results shown are representative of four experiments.

Techniques Used: Flow Cytometry, Cell Culture, Cytometry, Staining

Time course of VCAM-1 expression in fibroblast-like synoviocytes treated with a single application of IL-4 ( A ) and IL-13 ( B ). Cells were cultured with either medium only ( open circles ), TNF-α (10 ng/ml) ( closed circles ), IL-4 (10 ng/ml) ( open squares ), IL-4 plus TNF-α (both at 10 ng/ml) ( closed squares ) , IL-13 (10 ng/ml) ( open triangles ), or IL-13 plus TNF-α (both at 10 ng/ml) ( closed triangles ). Cell surface VCAM-1 expression was analyzed at days 1, 3, 7, and 14 by flow cytometry using mAb BBA-5. Samples were analyzed using a FACScan and the results expressed as corrected MFI. Data are presented as the mean ± SE of four or five experiments. *, P
Figure Legend Snippet: Time course of VCAM-1 expression in fibroblast-like synoviocytes treated with a single application of IL-4 ( A ) and IL-13 ( B ). Cells were cultured with either medium only ( open circles ), TNF-α (10 ng/ml) ( closed circles ), IL-4 (10 ng/ml) ( open squares ), IL-4 plus TNF-α (both at 10 ng/ml) ( closed squares ) , IL-13 (10 ng/ml) ( open triangles ), or IL-13 plus TNF-α (both at 10 ng/ml) ( closed triangles ). Cell surface VCAM-1 expression was analyzed at days 1, 3, 7, and 14 by flow cytometry using mAb BBA-5. Samples were analyzed using a FACScan and the results expressed as corrected MFI. Data are presented as the mean ± SE of four or five experiments. *, P

Techniques Used: Expressing, Cell Culture, Flow Cytometry, Cytometry

Chronic-cytokine treatment with TNF-α and IL-1β prolongs transient VCAM-1 expression on fibroblast-like synoviocytes. Medium plus IL-1β (10 ng/ml) ( closed circles ), medium plus TNF-α (10 ng/ml), or medium alone ( open circles ) were added at day 0 and re-added at days 3, 6, and 9, over a 12-day period. Cell surface VCAM-1 expression was determined 48 hours after each cytokine addition using flow cytometry and mAb BBA-5 as primary antibody. Samples were analyzed using a FACScan and the results expressed as corrected MFI. Data are presented as the mean ± SE of six experiments. *, P
Figure Legend Snippet: Chronic-cytokine treatment with TNF-α and IL-1β prolongs transient VCAM-1 expression on fibroblast-like synoviocytes. Medium plus IL-1β (10 ng/ml) ( closed circles ), medium plus TNF-α (10 ng/ml), or medium alone ( open circles ) were added at day 0 and re-added at days 3, 6, and 9, over a 12-day period. Cell surface VCAM-1 expression was determined 48 hours after each cytokine addition using flow cytometry and mAb BBA-5 as primary antibody. Samples were analyzed using a FACScan and the results expressed as corrected MFI. Data are presented as the mean ± SE of six experiments. *, P

Techniques Used: Expressing, Flow Cytometry, Cytometry

IL-1β and TNF-α induce transient VCAM-1 expression on fibroblast-like synoviocytes. Cells were treated with either medium only ( open circles ) , or medium plus IL-1β (10 ng/ml) ( closed circles ), or medium plus TNF-α (10 ng/ml) ( open squares ) for 1, 2, 3, or 7 days. VCAM-1 cell-surface expression was determined by flow cytometry using mAb BBA-5 as primary antibody. Samples were analyzed using a FACScan and the results expressed as corrected MFI. Data are presented as the mean ± SE of six experiments. *, P
Figure Legend Snippet: IL-1β and TNF-α induce transient VCAM-1 expression on fibroblast-like synoviocytes. Cells were treated with either medium only ( open circles ) , or medium plus IL-1β (10 ng/ml) ( closed circles ), or medium plus TNF-α (10 ng/ml) ( open squares ) for 1, 2, 3, or 7 days. VCAM-1 cell-surface expression was determined by flow cytometry using mAb BBA-5 as primary antibody. Samples were analyzed using a FACScan and the results expressed as corrected MFI. Data are presented as the mean ± SE of six experiments. *, P

Techniques Used: Expressing, Flow Cytometry, Cytometry

8) Product Images from "MD41, a novel T helper 0 clone, mediates mast-cell dependent delayed-type hypersensitivity in mice"

Article Title: MD41, a novel T helper 0 clone, mediates mast-cell dependent delayed-type hypersensitivity in mice

Journal: Immunology

doi: 10.1046/j.1365-2567.2002.01509.x

MD41 cells simultaneously produce IL-4 and IFN-γ at single cell level after the Con A stimulation. MD41 cells cultured with Con A or medium alone for 24 hr were fixed in 4% paraformaldehyde and washed with HBSS supplemented with 0·5% saponin. Cells were stained with rat anti IL-4 and FITC-labelled anti-rat immunoglobulin as described in Materials and Methods. After two washes with HBSS containing 0·5% saponin, cells were stained with PE conjugated anti-IFN-γ. After 20 min, cells were washed twice with PBS/BSA/saponin and then with PBS/BSA without saponin to allow membrane closure. Samples were analysed on a FACScan flow cytometer. Frequencies of IFN-γ-producing cells are shown on the y -axis and IL-4-producing cells on the x -axis. Staining with anti-IFN-γ-PE alone (a), staining with anti-IL-4 alone (b), and double staining (c).
Figure Legend Snippet: MD41 cells simultaneously produce IL-4 and IFN-γ at single cell level after the Con A stimulation. MD41 cells cultured with Con A or medium alone for 24 hr were fixed in 4% paraformaldehyde and washed with HBSS supplemented with 0·5% saponin. Cells were stained with rat anti IL-4 and FITC-labelled anti-rat immunoglobulin as described in Materials and Methods. After two washes with HBSS containing 0·5% saponin, cells were stained with PE conjugated anti-IFN-γ. After 20 min, cells were washed twice with PBS/BSA/saponin and then with PBS/BSA without saponin to allow membrane closure. Samples were analysed on a FACScan flow cytometer. Frequencies of IFN-γ-producing cells are shown on the y -axis and IL-4-producing cells on the x -axis. Staining with anti-IFN-γ-PE alone (a), staining with anti-IL-4 alone (b), and double staining (c).

Techniques Used: Cell Culture, Staining, Flow Cytometry, Cytometry, Double Staining

Phenotypic characterization of the MD41 clone established from lymph node cells of mice sensitized with MHSA in IFA. MD41 cells were stained with monoclonal antibodies against the lymphocyte markers CD90·2 (a), CD4 (b), CD8 (c), and CD45R (d). After two washes, the cells were analysed by flow cytometry. Fluorescence intensity of background staining (…) or specific binding in the presence of primary mAbs (—) was determined using a FACScan.
Figure Legend Snippet: Phenotypic characterization of the MD41 clone established from lymph node cells of mice sensitized with MHSA in IFA. MD41 cells were stained with monoclonal antibodies against the lymphocyte markers CD90·2 (a), CD4 (b), CD8 (c), and CD45R (d). After two washes, the cells were analysed by flow cytometry. Fluorescence intensity of background staining (…) or specific binding in the presence of primary mAbs (—) was determined using a FACScan.

Techniques Used: Mouse Assay, Immunofluorescence, Staining, Flow Cytometry, Cytometry, Fluorescence, Binding Assay

9) Product Images from "Novel compound cedrelone inhibits hepatocellular carcinoma progression via PBLD and Ras/Rap1"

Article Title: Novel compound cedrelone inhibits hepatocellular carcinoma progression via PBLD and Ras/Rap1

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2019.8080

PBLD is a key target in cedrelone treated HCC. (A) Activation effect of cedrelone on PBLD expression in HepG2 and Hep3B cells was detected via western blotting after cells were treated with 20 µM cedrelone for 24 h. HepG2 and Hep3B cells were transfected with PBLD siRNA, which knocked down PBLD expression. HepG2 and Hep3B cells were treated with 20 µM cedrelone for 24 h. The effects of cedrelone on (B) PBLD expression, (C) cell growth, (D) cell viability and (E) cell death were detected in HepG2 and Hep3B cells via (B) western blotting, (C) sulforhodamine B staining, (D) a Cell Counting kit-8 assay and (E) trypan blue staining. HepG2 and Hep3B cells were transfected with PBLD siRNA and treated with 20 µM cedrelone for 24 h. (F) detection of cell invasion and (G) statistical analysis and (H) detection of cell migration and (I) statistical analysis. Scale bar, 200 µm for all eight images. HepG2 and Hep3B cells were transfected with PBLD siRNA or con-siRNA and treated with 20 µM cedrelone for 24 h. (J) epithelial-mesenchymal transition markers were detected through western blot. (K) The cell apoptosis were detected through FACScan flow cytometry, the abscissa represents Annexin V staining whilst the ordinate represents propidium iodide staining, and (L) statistical analysis. (M) The apoptosis markers were detected using western blot analysis. *P
Figure Legend Snippet: PBLD is a key target in cedrelone treated HCC. (A) Activation effect of cedrelone on PBLD expression in HepG2 and Hep3B cells was detected via western blotting after cells were treated with 20 µM cedrelone for 24 h. HepG2 and Hep3B cells were transfected with PBLD siRNA, which knocked down PBLD expression. HepG2 and Hep3B cells were treated with 20 µM cedrelone for 24 h. The effects of cedrelone on (B) PBLD expression, (C) cell growth, (D) cell viability and (E) cell death were detected in HepG2 and Hep3B cells via (B) western blotting, (C) sulforhodamine B staining, (D) a Cell Counting kit-8 assay and (E) trypan blue staining. HepG2 and Hep3B cells were transfected with PBLD siRNA and treated with 20 µM cedrelone for 24 h. (F) detection of cell invasion and (G) statistical analysis and (H) detection of cell migration and (I) statistical analysis. Scale bar, 200 µm for all eight images. HepG2 and Hep3B cells were transfected with PBLD siRNA or con-siRNA and treated with 20 µM cedrelone for 24 h. (J) epithelial-mesenchymal transition markers were detected through western blot. (K) The cell apoptosis were detected through FACScan flow cytometry, the abscissa represents Annexin V staining whilst the ordinate represents propidium iodide staining, and (L) statistical analysis. (M) The apoptosis markers were detected using western blot analysis. *P

Techniques Used: Activation Assay, Expressing, Western Blot, Transfection, Staining, Cell Counting, Migration, Flow Cytometry, Cytometry

Cedrelone mediates cell bioactivities in hepatocellular carcinoma. (A) Chemical structure of cedrelone. (B) Cells were treated with different concentrations of cedrelone for 24 h and the growth of Hep3B and HepG2 cells was detected via sulforhodamine B staining. (C) Cells were treated with different concentrations of cedrelone for 24 h and a Cell Counting kit-8 assay was used to measure cell viability. (D) Cells were treated with different concentrations of cedrelone for 24 h and trypan blue staining was used to detect the cell death ratio. Cells were treated with 20 µM cedrelone for 24 h and cell, (E) detection of cell invasion and (F) statistical analysis, (G) detection the cell migration and (H) statistical analysis. Scale bar, 200 µm. (I) Epithelial-mesenchymal transition markers were detected via western blotting after cells were treated with 20 µM cedrelone for 24 h. (J) The cell apoptosis ratio was detected using a FACScan flow cytometer, the abscissa represents the level of Annexin V staining whilst the ordinate represents propidium iodide staining. (K) Quantified results from (J). (L) Apoptosis marker levels were detected via western blotting after cells were treated with 20 µM cedrelone for 24 h. In figs B, C and D, *P
Figure Legend Snippet: Cedrelone mediates cell bioactivities in hepatocellular carcinoma. (A) Chemical structure of cedrelone. (B) Cells were treated with different concentrations of cedrelone for 24 h and the growth of Hep3B and HepG2 cells was detected via sulforhodamine B staining. (C) Cells were treated with different concentrations of cedrelone for 24 h and a Cell Counting kit-8 assay was used to measure cell viability. (D) Cells were treated with different concentrations of cedrelone for 24 h and trypan blue staining was used to detect the cell death ratio. Cells were treated with 20 µM cedrelone for 24 h and cell, (E) detection of cell invasion and (F) statistical analysis, (G) detection the cell migration and (H) statistical analysis. Scale bar, 200 µm. (I) Epithelial-mesenchymal transition markers were detected via western blotting after cells were treated with 20 µM cedrelone for 24 h. (J) The cell apoptosis ratio was detected using a FACScan flow cytometer, the abscissa represents the level of Annexin V staining whilst the ordinate represents propidium iodide staining. (K) Quantified results from (J). (L) Apoptosis marker levels were detected via western blotting after cells were treated with 20 µM cedrelone for 24 h. In figs B, C and D, *P

Techniques Used: Staining, Cell Counting, Migration, Western Blot, Flow Cytometry, Cytometry, Marker

10) Product Images from "Kinetics of GPIbα-vWF-A1 Tether Bond under Flow: Effect of GPIbα Mutations on the Association and Dissociation Rates"

Article Title: Kinetics of GPIbα-vWF-A1 Tether Bond under Flow: Effect of GPIbα Mutations on the Association and Dissociation Rates

Journal: Biophysical Journal

doi:

( A ) Cell surface expression of GPIb α (WT, Q232V, and K237V) on transfected CHO cells: The cells were incubated with FITC-conjugated mouse monoclonal antibody AK2 in 5% HSA solution for 60 min. The expression levels were detected using FACScan flow cytometry. CHO cells expressing GPIb β -IX (without α -subunit) were used as control. ( B ) Adsorption of VWF-A1 on glass coverslips at 37°C. The absorbance values are averaged from two experiments performed in triplicate.
Figure Legend Snippet: ( A ) Cell surface expression of GPIb α (WT, Q232V, and K237V) on transfected CHO cells: The cells were incubated with FITC-conjugated mouse monoclonal antibody AK2 in 5% HSA solution for 60 min. The expression levels were detected using FACScan flow cytometry. CHO cells expressing GPIb β -IX (without α -subunit) were used as control. ( B ) Adsorption of VWF-A1 on glass coverslips at 37°C. The absorbance values are averaged from two experiments performed in triplicate.

Techniques Used: Expressing, Transfection, Incubation, Flow Cytometry, Cytometry, Adsorption

11) Product Images from "Anti-Cancer Effects of Protein Extracts from Calvatia lilacina, Pleurotus ostreatus and Volvariella volvacea"

Article Title: Anti-Cancer Effects of Protein Extracts from Calvatia lilacina, Pleurotus ostreatus and Volvariella volvacea

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1093/ecam/neq057

Intracellular ROS evaluation of PO and VV protein extract-treated SW480 cells. SW480 cells were exposed to PO and VV protein extracts (100 μ g mL −1 ) for 24 h. Production of intracellular ROS was detected by flow cytometry using DCFH-DA. The intracellular fluorescence of DCF was measured with a Becton-Dickinson FACScan flow cytometer. Data in each panel represent the DCF fluorescence intensity within the cells. The values shown are the means ± SD ( n = 5–8 of individual experiments). * P
Figure Legend Snippet: Intracellular ROS evaluation of PO and VV protein extract-treated SW480 cells. SW480 cells were exposed to PO and VV protein extracts (100 μ g mL −1 ) for 24 h. Production of intracellular ROS was detected by flow cytometry using DCFH-DA. The intracellular fluorescence of DCF was measured with a Becton-Dickinson FACScan flow cytometer. Data in each panel represent the DCF fluorescence intensity within the cells. The values shown are the means ± SD ( n = 5–8 of individual experiments). * P

Techniques Used: Flow Cytometry, Cytometry, Fluorescence

12) Product Images from "Augmented efficacy of brentuximab vedotin combined with ruxolitinib and/or Navitoclax in a murine model of human Hodgkin’s lymphoma"

Article Title: Augmented efficacy of brentuximab vedotin combined with ruxolitinib and/or Navitoclax in a murine model of human Hodgkin’s lymphoma

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1524668113

The combination of ruxolitinib, Navitoclax, and BV triggers a mitochondrial apoptosis pathway in HDLM-2 cells. The mitochondrial membrane potential was measured by the membrane-permeant JC-1 dye staining. After treatment (ruxolitinib, 250 nM; Navitoclax, 50 nM; and BV, 100 ng/mL) for 48 h, the HDLM-2 cells were incubated with JC-1 at a concentration of 10 µg/mL in medium at 37 °C, 5% CO 2 for 10 min and then washed with PBS. The samples were collected on a FACScan flow cytometer. Data are representative of three independent experiments.
Figure Legend Snippet: The combination of ruxolitinib, Navitoclax, and BV triggers a mitochondrial apoptosis pathway in HDLM-2 cells. The mitochondrial membrane potential was measured by the membrane-permeant JC-1 dye staining. After treatment (ruxolitinib, 250 nM; Navitoclax, 50 nM; and BV, 100 ng/mL) for 48 h, the HDLM-2 cells were incubated with JC-1 at a concentration of 10 µg/mL in medium at 37 °C, 5% CO 2 for 10 min and then washed with PBS. The samples were collected on a FACScan flow cytometer. Data are representative of three independent experiments.

Techniques Used: Staining, Incubation, Concentration Assay, Flow Cytometry, Cytometry

13) Product Images from "Reducing AD-Like Pathology in 3xTg-AD Mouse Model by DNA Epitope Vaccine -- A Novel Immunotherapeutic Strategy"

Article Title: Reducing AD-Like Pathology in 3xTg-AD Mouse Model by DNA Epitope Vaccine -- A Novel Immunotherapeutic Strategy

Journal: PLoS ONE

doi: 10.1371/journal.pone.0002124

Schematic representation of DNA vaccine construct, its expression, and immunizations schedules. A) Fusion gene encoding epitope vaccine, 3Aβ 1–11 and PADRE (aKXVAAWTLKAAaZC, x = L-cyclohexylalanine, Z = aminocaproic acid) was introduced into pCMVE-AB/hMDC in frame to the 3′-end of mature MDC exactly as shown. MDC gene is fused to the IP10 signal sequence in its 5′-end and to the 3Aβ 1–11 -PADRE minigene in its 3′-end through small spacer. Transcription of MDC-3Aβ 1–11 -PADRE gene is controlled by CMV promoter/enhancer. B) To analyze the intracellular expression of pMDC-3Aβ 1–11 -PADRE, transiently transfected CHO cells were permeabilized/fixed and stained with anti-Aβ 6E10 antibody followed by FITC-conjugated anti mouse IgG. Cells were analyzed by FACScan. Approximately ∼65% of CHO cells transfected with pMDC-3Aβ 1–11 -PADRE showed 6E10-positive staining. Cells transfected with control vector showed only background staining. C) The secretion of MDC-3Aβ 1–11 -PADRE by transfected CHO cells was demonstrated by IP/WB. MDC-3Aβ 1–11 -PADRE protein was immunoprecipitated from the growth medium of transfected CHO cells with anti-Aβ antibody (6E10), separated by 15% Tris-SDS PAGE and transferred onto nitrocellulose membrane. Proteins were visualized by staining with 6E10 followed by HRP-conjugated anti-mouse IgG. Lane 1 represent vector (pCMVE-AB/MDC); Lane 2 represent Epitope Vaccine (pMDC-3Aβ 1–11 -PADRE). D) 3–4 mo old C57/Bl6 (n = 6) or 3xTg-AD (n = 4) mice were immunized 5 times biweekly. Blood was collected after 3 rd , 4 th and 5 th immunizations and humoral immune response was analyzed in sera. Seven days after the last immunization mice were sacrificed and cellular immune response was analyzed in splenocytes. Experiment with C57/Bl6 mice was repeated (total n = 12). Next, 3–4 mo old 3xTg-AD (n = 9) mice were immunized 11 times as indicated. Blood was collected and humoral immune response was analyzed in sera. After 11 th immunization behavioral studies were conducted and upon its completion, mice were sacrificed and neuropathological changes were analyzed in the brains of experimental and control mice.
Figure Legend Snippet: Schematic representation of DNA vaccine construct, its expression, and immunizations schedules. A) Fusion gene encoding epitope vaccine, 3Aβ 1–11 and PADRE (aKXVAAWTLKAAaZC, x = L-cyclohexylalanine, Z = aminocaproic acid) was introduced into pCMVE-AB/hMDC in frame to the 3′-end of mature MDC exactly as shown. MDC gene is fused to the IP10 signal sequence in its 5′-end and to the 3Aβ 1–11 -PADRE minigene in its 3′-end through small spacer. Transcription of MDC-3Aβ 1–11 -PADRE gene is controlled by CMV promoter/enhancer. B) To analyze the intracellular expression of pMDC-3Aβ 1–11 -PADRE, transiently transfected CHO cells were permeabilized/fixed and stained with anti-Aβ 6E10 antibody followed by FITC-conjugated anti mouse IgG. Cells were analyzed by FACScan. Approximately ∼65% of CHO cells transfected with pMDC-3Aβ 1–11 -PADRE showed 6E10-positive staining. Cells transfected with control vector showed only background staining. C) The secretion of MDC-3Aβ 1–11 -PADRE by transfected CHO cells was demonstrated by IP/WB. MDC-3Aβ 1–11 -PADRE protein was immunoprecipitated from the growth medium of transfected CHO cells with anti-Aβ antibody (6E10), separated by 15% Tris-SDS PAGE and transferred onto nitrocellulose membrane. Proteins were visualized by staining with 6E10 followed by HRP-conjugated anti-mouse IgG. Lane 1 represent vector (pCMVE-AB/MDC); Lane 2 represent Epitope Vaccine (pMDC-3Aβ 1–11 -PADRE). D) 3–4 mo old C57/Bl6 (n = 6) or 3xTg-AD (n = 4) mice were immunized 5 times biweekly. Blood was collected after 3 rd , 4 th and 5 th immunizations and humoral immune response was analyzed in sera. Seven days after the last immunization mice were sacrificed and cellular immune response was analyzed in splenocytes. Experiment with C57/Bl6 mice was repeated (total n = 12). Next, 3–4 mo old 3xTg-AD (n = 9) mice were immunized 11 times as indicated. Blood was collected and humoral immune response was analyzed in sera. After 11 th immunization behavioral studies were conducted and upon its completion, mice were sacrificed and neuropathological changes were analyzed in the brains of experimental and control mice.

Techniques Used: Construct, Expressing, Sequencing, Transfection, Staining, Plasmid Preparation, Western Blot, Immunoprecipitation, SDS Page, Mouse Assay

14) Product Images from "PERK Signal-Modulated Protein Translation Promotes the Survivability of Dengue 2 Virus-Infected Mosquito Cells and Extends Viral Replication"

Article Title: PERK Signal-Modulated Protein Translation Promotes the Survivability of Dengue 2 Virus-Infected Mosquito Cells and Extends Viral Replication

Journal: Viruses

doi: 10.3390/v9090262

Evaluation of endoplasmic reticular (ER) stress by measuring the mitochondrial membrane potential (MMP) in dengue 2 virus (DENV2)-infected C6/36 cells. To measure MMP, detection of the FITC channel for green MitoCapture monomers by FACScan flow cytometry was carried out in DENV2-infected C6/36 cells (MOI = 1) with and without PERK inhibitor (5 μM) treatment. The MMP in the group treated with the PERK inhibitor had significantly increased at 48 h post-infection (hpi) (a 2.32-fold increase) compared to cells in the two control groups (1.20-fold for the untreated and 1.36-fold increases for the DMSO-treated group) (Student’s t -test; *** p
Figure Legend Snippet: Evaluation of endoplasmic reticular (ER) stress by measuring the mitochondrial membrane potential (MMP) in dengue 2 virus (DENV2)-infected C6/36 cells. To measure MMP, detection of the FITC channel for green MitoCapture monomers by FACScan flow cytometry was carried out in DENV2-infected C6/36 cells (MOI = 1) with and without PERK inhibitor (5 μM) treatment. The MMP in the group treated with the PERK inhibitor had significantly increased at 48 h post-infection (hpi) (a 2.32-fold increase) compared to cells in the two control groups (1.20-fold for the untreated and 1.36-fold increases for the DMSO-treated group) (Student’s t -test; *** p

Techniques Used: Infection, Flow Cytometry, Cytometry

The PERK signaling pathway is involved in generating reactive oxygen species (ROS) accumulation in C6/36 cells with dengue 2 virus (DENV2) infection. Superoxide anions (O 2− ) are one of the main ROS and are usually converted to hydrogen peroxide (H 2 O 2 ). ( A ) Using FACScan flow cytometry to detect the relative fluorescent intensity, superoxide anions exhibited no obvious change in cells with DENV2 infection (MOI = 1) at 24 h post-infection (hpi; 1.34-fold) even though a PERK inhibitor (5 μM) was applied. A significant (1.82-fold) increase in superoxide anions was measured at 48 hpi, compared to those in the control groups (1.07-fold for the untreated and 1.16-fold for the DMSO-treated group; Student’s t -test; *** p
Figure Legend Snippet: The PERK signaling pathway is involved in generating reactive oxygen species (ROS) accumulation in C6/36 cells with dengue 2 virus (DENV2) infection. Superoxide anions (O 2− ) are one of the main ROS and are usually converted to hydrogen peroxide (H 2 O 2 ). ( A ) Using FACScan flow cytometry to detect the relative fluorescent intensity, superoxide anions exhibited no obvious change in cells with DENV2 infection (MOI = 1) at 24 h post-infection (hpi; 1.34-fold) even though a PERK inhibitor (5 μM) was applied. A significant (1.82-fold) increase in superoxide anions was measured at 48 hpi, compared to those in the control groups (1.07-fold for the untreated and 1.16-fold for the DMSO-treated group; Student’s t -test; *** p

Techniques Used: Infection, Flow Cytometry, Cytometry

The effect of the PERK inhibitor on activation of caspases-9 (the initiator) and -3 (the effector) in C6/36 cells with dengue 2 virus (DENV2) infection by using FACScan flow cytometry. ( A ) In DENV2-infected C6/36 cells (MOI = 1), no evident change in caspase-9 had occurred in cells by 24 h post-infection (hpi), even when they were treated with a PERK inhibitor (5 μM). However, it had increased by 48 hpi (2.52-fold increase) compared to the two control groups (1.20- and 1.18-fold for untreated and DMSO-treated cells, respectively; Student’s t -test; *** p
Figure Legend Snippet: The effect of the PERK inhibitor on activation of caspases-9 (the initiator) and -3 (the effector) in C6/36 cells with dengue 2 virus (DENV2) infection by using FACScan flow cytometry. ( A ) In DENV2-infected C6/36 cells (MOI = 1), no evident change in caspase-9 had occurred in cells by 24 h post-infection (hpi), even when they were treated with a PERK inhibitor (5 μM). However, it had increased by 48 hpi (2.52-fold increase) compared to the two control groups (1.20- and 1.18-fold for untreated and DMSO-treated cells, respectively; Student’s t -test; *** p

Techniques Used: Activation Assay, Infection, Flow Cytometry, Cytometry

15) Product Images from "MAPK signaling mediates sinomenine hydrochloride-induced human breast cancer cell death via both reactive oxygen species-dependent and -independent pathways: an in vitro and in vivo study"

Article Title: MAPK signaling mediates sinomenine hydrochloride-induced human breast cancer cell death via both reactive oxygen species-dependent and -independent pathways: an in vitro and in vivo study

Journal: Cell Death & Disease

doi: 10.1038/cddis.2014.321

SH triggered ROS generation in breast cancer cells. ( a and b ) DCFH-DA staining was used to detect ROS production in MDA-MB-231 and MCF-7. ( c ) Pretreatment of 5 mM/l NAC for 2 h significantly abolished ROS generation induced by SH. Cells were treated with 1.0 μ mol/ml SH and 5 mM/l NAC, independently or in combination, and ROS level at 48 h was analyzed using a FACSCAN flow cytometer. Data are the means±S.D. of three independent experiments. * P
Figure Legend Snippet: SH triggered ROS generation in breast cancer cells. ( a and b ) DCFH-DA staining was used to detect ROS production in MDA-MB-231 and MCF-7. ( c ) Pretreatment of 5 mM/l NAC for 2 h significantly abolished ROS generation induced by SH. Cells were treated with 1.0 μ mol/ml SH and 5 mM/l NAC, independently or in combination, and ROS level at 48 h was analyzed using a FACSCAN flow cytometer. Data are the means±S.D. of three independent experiments. * P

Techniques Used: Staining, Multiple Displacement Amplification, Flow Cytometry, Cytometry

SH induced G1/S cell cycle arrest in human breast cancer cells. ( a and b ) Cells were treated with SH for 48 h and subjected to DNA content analysis using a FACSCAN flow cytometer. ( c ) Proteins involved in G1/S transition were analyzed by western blotting. Cells were treated with SH (0, 0.25, 0.5 and 1.0 μ mol/ml) for 48 h, and total proteins were extracted. Equal protein loading was evaluated by β -actin. Data are represented as mean±S.D. of three independent experiments. * P
Figure Legend Snippet: SH induced G1/S cell cycle arrest in human breast cancer cells. ( a and b ) Cells were treated with SH for 48 h and subjected to DNA content analysis using a FACSCAN flow cytometer. ( c ) Proteins involved in G1/S transition were analyzed by western blotting. Cells were treated with SH (0, 0.25, 0.5 and 1.0 μ mol/ml) for 48 h, and total proteins were extracted. Equal protein loading was evaluated by β -actin. Data are represented as mean±S.D. of three independent experiments. * P

Techniques Used: Flow Cytometry, Cytometry, Western Blot

16) Product Images from "Polydatin, a natural precursor of resveratrol, induces cell cycle arrest and differentiation of human colorectal Caco-2 cell"

Article Title: Polydatin, a natural precursor of resveratrol, induces cell cycle arrest and differentiation of human colorectal Caco-2 cell

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-264

Effect of ISBn alone or in combination on mithocondrial superoxide anions, nitric oxide, and enzyme scavengers activity in Caco-2 cells. (A) After 24 h Caco-2 treatment with polydatin and resveratrol alone or in combination (240 and 100 μM) at 37°C, the mitochondrial superoxide anion production was analysed by HE (20 ng · mL-1) staining. Dye accumulation was analysed by FACScan flow cytometer (FACScan, Becton Dickinson) by the CellQuest software, and the intensities of the bands were expressed as percent control. For each sample, 2 × 10 4 events were acquired. (B) Nitric oxide was measured in medium. Whole cell homogenates were used for evaluated the Catalase (C) and Mn-SOD activity (D) . The bars represent means ± SEM of three independent experiments. Asterisks indicate significant difference between the Caco-2-treated samples compared with control value ** P
Figure Legend Snippet: Effect of ISBn alone or in combination on mithocondrial superoxide anions, nitric oxide, and enzyme scavengers activity in Caco-2 cells. (A) After 24 h Caco-2 treatment with polydatin and resveratrol alone or in combination (240 and 100 μM) at 37°C, the mitochondrial superoxide anion production was analysed by HE (20 ng · mL-1) staining. Dye accumulation was analysed by FACScan flow cytometer (FACScan, Becton Dickinson) by the CellQuest software, and the intensities of the bands were expressed as percent control. For each sample, 2 × 10 4 events were acquired. (B) Nitric oxide was measured in medium. Whole cell homogenates were used for evaluated the Catalase (C) and Mn-SOD activity (D) . The bars represent means ± SEM of three independent experiments. Asterisks indicate significant difference between the Caco-2-treated samples compared with control value ** P

Techniques Used: Activity Assay, Staining, Flow Cytometry, Cytometry, Software

17) Product Images from "PGC-1-Related Coactivator Modulates Mitochondrial-Nuclear Crosstalk through Endogenous Nitric Oxide in a Cellular Model of Oncocytic Thyroid Tumours"

Article Title: PGC-1-Related Coactivator Modulates Mitochondrial-Nuclear Crosstalk through Endogenous Nitric Oxide in a Cellular Model of Oncocytic Thyroid Tumours

Journal: PLoS ONE

doi: 10.1371/journal.pone.0007964

Effect of 100 µM SNAP treatment during 96 h on XTC.UC1 and BCPAP cell lines ( N = 5 per cell line). 1A: Nitric oxide measurement with the FACScan cytometer (10 µM final DAF2/DA dye). Results are expressed in arbitrary fluorescent units as mean values±SEM. * P ≤0.05 versus T0. ** P ≤0.05 versus T0 and T72. 1B: Evaluation of protein nitration by Western blotting using antibody raised against nitrotyrosine-modified proteins. Immunoblots are quantified by densitometric analysis and expressed in arbitrary units (relative to α-tubulin) as mean values±SEM. * P ≤0.05 versus T0.
Figure Legend Snippet: Effect of 100 µM SNAP treatment during 96 h on XTC.UC1 and BCPAP cell lines ( N = 5 per cell line). 1A: Nitric oxide measurement with the FACScan cytometer (10 µM final DAF2/DA dye). Results are expressed in arbitrary fluorescent units as mean values±SEM. * P ≤0.05 versus T0. ** P ≤0.05 versus T0 and T72. 1B: Evaluation of protein nitration by Western blotting using antibody raised against nitrotyrosine-modified proteins. Immunoblots are quantified by densitometric analysis and expressed in arbitrary units (relative to α-tubulin) as mean values±SEM. * P ≤0.05 versus T0.

Techniques Used: Cytometry, Nitration, Western Blot, Modification

18) Product Images from "A modified p53 enhances apoptosis in sarcoma cell lines mediated by doxorubicin"

Article Title: A modified p53 enhances apoptosis in sarcoma cell lines mediated by doxorubicin

Journal: British Journal of Cancer

doi: 10.1038/sj.bjc.6601653

Ad- p53 14/19 enhances doxorubicin-mediated apoptosis in SJSA osteosarcoma cells. Cells were infected with NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 for 3 days alone or in combination with 0.1 μ g ml −1 doxorubicin. Cells were then stained with PI and apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer. Examples of apoptosis in SJSA cells are shown in ( A ). ( B ) Percentages of apoptotic cells are given as the mean+standard deviation of Log [PI] from three independent experiments. * Indicates a significant increase compared to Ad-wt p 53 plus doxorubicin treatment ( P
Figure Legend Snippet: Ad- p53 14/19 enhances doxorubicin-mediated apoptosis in SJSA osteosarcoma cells. Cells were infected with NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 for 3 days alone or in combination with 0.1 μ g ml −1 doxorubicin. Cells were then stained with PI and apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer. Examples of apoptosis in SJSA cells are shown in ( A ). ( B ) Percentages of apoptotic cells are given as the mean+standard deviation of Log [PI] from three independent experiments. * Indicates a significant increase compared to Ad-wt p 53 plus doxorubicin treatment ( P

Techniques Used: Infection, Staining, Flow Cytometry, Cytometry, Standard Deviation

Ad- p53 14/19 combined with cisplatin induces more significant apoptosis than Ad-wt p53 combined with cisplatin in SJSA cells. Cells were infected with NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 for 3 days alone or in combination with 5 μg/ml cisplatin. Cells were then stained with PI and apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer. Percentages of apoptosis in HT1080 cells are shown in ( A ), and percentages of apoptosis in SJSA cells are shown in ( B ). Values shown are the mean+standard deviation of Log [PI] from three independent experiments. * Indicates a significant increase compared to Ad-wt p 53 plus cisplatin treatment ( P
Figure Legend Snippet: Ad- p53 14/19 combined with cisplatin induces more significant apoptosis than Ad-wt p53 combined with cisplatin in SJSA cells. Cells were infected with NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 for 3 days alone or in combination with 5 μg/ml cisplatin. Cells were then stained with PI and apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer. Percentages of apoptosis in HT1080 cells are shown in ( A ), and percentages of apoptosis in SJSA cells are shown in ( B ). Values shown are the mean+standard deviation of Log [PI] from three independent experiments. * Indicates a significant increase compared to Ad-wt p 53 plus cisplatin treatment ( P

Techniques Used: Infection, Staining, Flow Cytometry, Cytometry, Standard Deviation

Ad- p53 14/19, Ad- p53 22/23 and Ad-wt p53 enhance doxorubicin (Doxo)-mediated apoptosis in HT1080 fibrosarcoma cells. Cells were infected with NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 for 3 days alone or in combination with 0.1 μ g/ml doxorubicin. Cells were then stained with PI and apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer. Examples of apoptosis are shown in ( A ). ( B ) Percentages of apoptotis are given as the mean+standard deviation of Log [PI] from three independent experiments. Gray bars indicate HT1080 cells treated by mock infection, doxorubicin, NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 alone. Black bars indicate cells treated with doxorubicin plus NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19.
Figure Legend Snippet: Ad- p53 14/19, Ad- p53 22/23 and Ad-wt p53 enhance doxorubicin (Doxo)-mediated apoptosis in HT1080 fibrosarcoma cells. Cells were infected with NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 for 3 days alone or in combination with 0.1 μ g/ml doxorubicin. Cells were then stained with PI and apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer. Examples of apoptosis are shown in ( A ). ( B ) Percentages of apoptotis are given as the mean+standard deviation of Log [PI] from three independent experiments. Gray bars indicate HT1080 cells treated by mock infection, doxorubicin, NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19 alone. Black bars indicate cells treated with doxorubicin plus NCV, Ad-wt p53 , Ad- p53 22/23 or Ad- p53 14/19.

Techniques Used: Infection, Staining, Flow Cytometry, Cytometry, Standard Deviation

Ad- p53 14/19 induces significant apoptosis in sarcoma cell lines. ( A ) HT1080 fibrosarcoma cells, ( B ) SJSA osteosarcoma cells and ( C ) NHF were infected with NCV or Ad-wt p53 , Ad- p53 14/19 or Ad- p53 22/23. Cells were stained with propidium iodide (PI), apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer and fold increase of apoptotic cells was calculated. Values shown are the mean+standard deviation of Log [PI] from three independent experiments. * Indicates a significant increase compare to Ad-wt p 53 treatment ( P
Figure Legend Snippet: Ad- p53 14/19 induces significant apoptosis in sarcoma cell lines. ( A ) HT1080 fibrosarcoma cells, ( B ) SJSA osteosarcoma cells and ( C ) NHF were infected with NCV or Ad-wt p53 , Ad- p53 14/19 or Ad- p53 22/23. Cells were stained with propidium iodide (PI), apoptosis was assessed with sub-G1 profile analysis using a FACScan flow cytometer and fold increase of apoptotic cells was calculated. Values shown are the mean+standard deviation of Log [PI] from three independent experiments. * Indicates a significant increase compare to Ad-wt p 53 treatment ( P

Techniques Used: Infection, Staining, Flow Cytometry, Cytometry, Standard Deviation

19) Product Images from "Recombinant TAT-BMI-1 fusion protein induces ex vivo expansion of human umbilical cord blood-derived hematopoietic stem cells"

Article Title: Recombinant TAT-BMI-1 fusion protein induces ex vivo expansion of human umbilical cord blood-derived hematopoietic stem cells

Journal: Oncotarget

doi: 10.18632/oncotarget.15156

Uptake of TAT-proteins by cord blood derived CD34 + and K562 cells ( A ) Purified TAT-GFP or TAT-BMI1 proteins were added to cultures of CD34 + cells at a final 100 nM concentration for 40 min at 37°C. Cytospins were fixed and TAT-proteins were detected by indirect immunofluorescence with rabbit anti-HA and anti-rabbit Alexafluor 488 antibodies as detailed in Materials and Methods; cell nuclei were counter-stained with DAPI. ( B ) Uptake of TAT-GFP and TAT-BMI-1 in CB-CD34 + cells after 40 and 120 minutes of incubation. Treatment and immunofluorescence were performed as described above, and the percentage of HA-positive/DAPI-positive cells was determined using ImageJ. ( C ) Subcellular localization of TAT-proteins. K562 cells (2 × 10 5 /ml) were incubated with 100 nM TAT-fusion proteins for 20 min. Cytoplasmic and nuclear extracts were prepared as described in the materials and methods section, and analyzed by western blotting using an anti-HA antibody to reveal the TAT-fusion proteins, as well as anti-HDAC1 and anti-actin as markers of the nuclear and cytosolic protein fractions respectively. As it can be observed, TAT-GFP accumulates in the cytosolic fraction and TAT-BMI-1 in the nuclear fraction. As controls, 10 ng of affinity-purified TAT-GFP (T-GFP) and TAT-BMI-1 (T-BMI1) were also loaded. ( D ) Accumulation kinetics of TAT-BMI-1. K562 cells were incubated with 100nM protein and collected after 30, 60, 120 and 240 min. After intracellular staining with anti-HA antibody and Alexafluor 488-conjugated secondary antibody, FACS analysis was performed on a BD FaCScan (Beckton-Dickinson, Milan, Italy). In the top part of the panel, blue histograms represent TAT-BMI-1-treated cells compared to (red histograms) untreated cells. The mean fluorescence intensity (MFI) values determined at each time point, minus the MFI of untreated cells, are shown in the graph in the lower part of the panel. ( E ) TAT-BMI-1 down-regulates INK4A expression. 1 × 10 5 CD34 + cells were stimulated for 3 days with repeated additions of 10 nM TAT-BMI-1. RNA was extracted and the INK4A and ARF mRNA levels were analyzed by RT-Q-PCR using GAPDH as a normalizing gene.
Figure Legend Snippet: Uptake of TAT-proteins by cord blood derived CD34 + and K562 cells ( A ) Purified TAT-GFP or TAT-BMI1 proteins were added to cultures of CD34 + cells at a final 100 nM concentration for 40 min at 37°C. Cytospins were fixed and TAT-proteins were detected by indirect immunofluorescence with rabbit anti-HA and anti-rabbit Alexafluor 488 antibodies as detailed in Materials and Methods; cell nuclei were counter-stained with DAPI. ( B ) Uptake of TAT-GFP and TAT-BMI-1 in CB-CD34 + cells after 40 and 120 minutes of incubation. Treatment and immunofluorescence were performed as described above, and the percentage of HA-positive/DAPI-positive cells was determined using ImageJ. ( C ) Subcellular localization of TAT-proteins. K562 cells (2 × 10 5 /ml) were incubated with 100 nM TAT-fusion proteins for 20 min. Cytoplasmic and nuclear extracts were prepared as described in the materials and methods section, and analyzed by western blotting using an anti-HA antibody to reveal the TAT-fusion proteins, as well as anti-HDAC1 and anti-actin as markers of the nuclear and cytosolic protein fractions respectively. As it can be observed, TAT-GFP accumulates in the cytosolic fraction and TAT-BMI-1 in the nuclear fraction. As controls, 10 ng of affinity-purified TAT-GFP (T-GFP) and TAT-BMI-1 (T-BMI1) were also loaded. ( D ) Accumulation kinetics of TAT-BMI-1. K562 cells were incubated with 100nM protein and collected after 30, 60, 120 and 240 min. After intracellular staining with anti-HA antibody and Alexafluor 488-conjugated secondary antibody, FACS analysis was performed on a BD FaCScan (Beckton-Dickinson, Milan, Italy). In the top part of the panel, blue histograms represent TAT-BMI-1-treated cells compared to (red histograms) untreated cells. The mean fluorescence intensity (MFI) values determined at each time point, minus the MFI of untreated cells, are shown in the graph in the lower part of the panel. ( E ) TAT-BMI-1 down-regulates INK4A expression. 1 × 10 5 CD34 + cells were stimulated for 3 days with repeated additions of 10 nM TAT-BMI-1. RNA was extracted and the INK4A and ARF mRNA levels were analyzed by RT-Q-PCR using GAPDH as a normalizing gene.

Techniques Used: Derivative Assay, Purification, Concentration Assay, Immunofluorescence, Staining, Incubation, Western Blot, Affinity Purification, FACS, Fluorescence, Expressing, Polymerase Chain Reaction

20) Product Images from "Genetic Evidence for an Interaction between Human Immunodeficiency Virus Type 1 Matrix and ?-Helix 2 of the gp41 Cytoplasmic Tail"

Article Title: Genetic Evidence for an Interaction between Human Immunodeficiency Virus Type 1 Matrix and ?-Helix 2 of the gp41 Cytoplasmic Tail

Journal: Journal of Virology

doi:

FACS analysis of cell surface Env expression. Cell surface immunostaining with an anti-gp41 monoclonal antibody (T32) was performed using CEM (12D-7) cells infected with wt or mutant NL4-3 virions pseudotyped with VSV-G. Averages of duplicate experiments are presented ± standard deviations. Solid bars represent mean fluorescence intensities determined by FACScan; open bars indicate percent Env-positive cells. Approximately 40% of cultures infected with the wt were Env positive.
Figure Legend Snippet: FACS analysis of cell surface Env expression. Cell surface immunostaining with an anti-gp41 monoclonal antibody (T32) was performed using CEM (12D-7) cells infected with wt or mutant NL4-3 virions pseudotyped with VSV-G. Averages of duplicate experiments are presented ± standard deviations. Solid bars represent mean fluorescence intensities determined by FACScan; open bars indicate percent Env-positive cells. Approximately 40% of cultures infected with the wt were Env positive.

Techniques Used: FACS, Expressing, Immunostaining, Infection, Mutagenesis, Fluorescence

21) Product Images from "Enhanced Gene Silencing in Cells Cured of Persistent Virus Infection by RNA Interference ▿"

Article Title: Enhanced Gene Silencing in Cells Cured of Persistent Virus Infection by RNA Interference ▿

Journal: Journal of Virology

doi: 10.1128/JVI.02060-09

Transfection efficiencies of fluorescein-conjugated siRNAs in HEp-2, HEp-Q4, and HEp-Q5 cells. A fluorescent siRNA-FITC (20 pmol) was used to transfect each of the three cell lines in the presence of Lipofectamine 2000. Fluorescent cells were analyzed 4 to 48 h posttransfection by using a FACScan flow cytometer (Becton Dickinson). The percentage of fluorescent cells (A) and the mean fluorescence per positive cell, in arbitrary units (B), are shown. Each bar represents the mean value ± SEM. (C) Representative FACS plots (cell granularity versus cell size), showing the similarities between the three cell populations.
Figure Legend Snippet: Transfection efficiencies of fluorescein-conjugated siRNAs in HEp-2, HEp-Q4, and HEp-Q5 cells. A fluorescent siRNA-FITC (20 pmol) was used to transfect each of the three cell lines in the presence of Lipofectamine 2000. Fluorescent cells were analyzed 4 to 48 h posttransfection by using a FACScan flow cytometer (Becton Dickinson). The percentage of fluorescent cells (A) and the mean fluorescence per positive cell, in arbitrary units (B), are shown. Each bar represents the mean value ± SEM. (C) Representative FACS plots (cell granularity versus cell size), showing the similarities between the three cell populations.

Techniques Used: Transfection, Flow Cytometry, Cytometry, Fluorescence, FACS

22) Product Images from "Cigarette smoke extract upregulates heme oxygenase-1 via PKC/NADPH oxidase/ROS/PDGFR/PI3K/Akt pathway in mouse brain endothelial cells"

Article Title: Cigarette smoke extract upregulates heme oxygenase-1 via PKC/NADPH oxidase/ROS/PDGFR/PI3K/Akt pathway in mouse brain endothelial cells

Journal: Journal of Neuroinflammation

doi: 10.1186/1742-2094-8-104

PPCSE-induced HO-1 expression is mediated via NADPH oxidase-dependent ROS generation in bEnd.3 cells . A: bEnd.3 cells were labeled with DCF-DA (10 μM), and then incubated with 30 μg/ml PPCSE for 30 min with/without pretreatment of NAC (3 mM), DPI (1 μM) or APO (1 μM) for 1 h. The fluorescence intensity (relative DCF fluorescence) was measured at 495 nm excitation and 529 nm emission using a FACScan flow cytometer. The NADPH oxidase activity was measured as described in the Methods. B: Cells were pretreated with APO for 1 h, and then incubated with PPCSE for 24 h. The expression of HO-1 was determined by Western blot. Data are summarized and expressed as the mean ± SEM of three individual experiments. * P
Figure Legend Snippet: PPCSE-induced HO-1 expression is mediated via NADPH oxidase-dependent ROS generation in bEnd.3 cells . A: bEnd.3 cells were labeled with DCF-DA (10 μM), and then incubated with 30 μg/ml PPCSE for 30 min with/without pretreatment of NAC (3 mM), DPI (1 μM) or APO (1 μM) for 1 h. The fluorescence intensity (relative DCF fluorescence) was measured at 495 nm excitation and 529 nm emission using a FACScan flow cytometer. The NADPH oxidase activity was measured as described in the Methods. B: Cells were pretreated with APO for 1 h, and then incubated with PPCSE for 24 h. The expression of HO-1 was determined by Western blot. Data are summarized and expressed as the mean ± SEM of three individual experiments. * P

Techniques Used: Expressing, Labeling, Incubation, Fluorescence, Flow Cytometry, Cytometry, Activity Assay, Western Blot

23) Product Images from "Reactive oxygen species contribute to oridonin-induced apoptosis and autophagy in human cervical carcinoma HeLa cells"

Article Title: Reactive oxygen species contribute to oridonin-induced apoptosis and autophagy in human cervical carcinoma HeLa cells

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2011.92

Significant reduction in mitochondrial transmembrane potential (ΔΨm) was induced by oridonin and could be rescued by NAC in HeLa cells. The cells were treated with 80 μmol/L oridonin for 24 h in the presence or absence of 5 mmol/L NAC and then were loaded with the membrane-sensitive probe rhodamine 123. The rhodamine retention was measured by flow cytometry. The corresponding linear diagram of the FACScan histograms is expressed at the base. n =3. Mean±SD. b P
Figure Legend Snippet: Significant reduction in mitochondrial transmembrane potential (ΔΨm) was induced by oridonin and could be rescued by NAC in HeLa cells. The cells were treated with 80 μmol/L oridonin for 24 h in the presence or absence of 5 mmol/L NAC and then were loaded with the membrane-sensitive probe rhodamine 123. The rhodamine retention was measured by flow cytometry. The corresponding linear diagram of the FACScan histograms is expressed at the base. n =3. Mean±SD. b P

Techniques Used: Flow Cytometry, Cytometry

ROS production was induced by oridonin and could be inhibited by NAC in HeLa cells. The cells were treated with various doses of oridonin or coincubated with 5 mmol/L NAC and 80 μmol/L oridonin for 24 h. The DCF-positive cells were measured by flow cytometry. The corresponding linear diagram of the FACScan histograms is expressed at the base. n =3. Mean±SD. c P
Figure Legend Snippet: ROS production was induced by oridonin and could be inhibited by NAC in HeLa cells. The cells were treated with various doses of oridonin or coincubated with 5 mmol/L NAC and 80 μmol/L oridonin for 24 h. The DCF-positive cells were measured by flow cytometry. The corresponding linear diagram of the FACScan histograms is expressed at the base. n =3. Mean±SD. c P

Techniques Used: Flow Cytometry, Cytometry

Persistent ROS generation was induced by oridonin in HeLa cells. The cells were treated with 80 μmol/L oridonin for 1, 3, 6, 12, and 24 h, and the DCF-positive cells were measured by flow cytometric analysis. The corresponding linear diagram of the FACScan histograms was expressed at the bottom. n =3. Mean±SD. c P
Figure Legend Snippet: Persistent ROS generation was induced by oridonin in HeLa cells. The cells were treated with 80 μmol/L oridonin for 1, 3, 6, 12, and 24 h, and the DCF-positive cells were measured by flow cytometric analysis. The corresponding linear diagram of the FACScan histograms was expressed at the bottom. n =3. Mean±SD. c P

Techniques Used: Flow Cytometry

24) Product Images from "Characterization of Human Mycobacterium bovis Bacille Calmette-Gu?rin-Reactive CD8+ T Cells"

Article Title: Characterization of Human Mycobacterium bovis Bacille Calmette-Gu?rin-Reactive CD8+ T Cells

Journal: Infection and Immunity

doi:

Flow cytometric detection of type 1 cytokine and perforin synthesis by CD8 + T cells. PBMC were stimulated for 6 days in the presence of live M. bovis BCG. CD8 + T cells were positively selected from the PBMC by the MACS separation system and incubated for 16 h in the presence of brefeldin A. Intracellular cytokine and perforin staining was performed before FACScan analysis.
Figure Legend Snippet: Flow cytometric detection of type 1 cytokine and perforin synthesis by CD8 + T cells. PBMC were stimulated for 6 days in the presence of live M. bovis BCG. CD8 + T cells were positively selected from the PBMC by the MACS separation system and incubated for 16 h in the presence of brefeldin A. Intracellular cytokine and perforin staining was performed before FACScan analysis.

Techniques Used: Flow Cytometry, Magnetic Cell Separation, Incubation, Staining

25) Product Images from "Production of Tumor Necrosis Factor Alpha in Human T Lymphocytes by Staphylococcal Enterotoxin B Correlates with Toxin-Induced Proliferation and Is Regulated through Protein Kinase C"

Article Title: Production of Tumor Necrosis Factor Alpha in Human T Lymphocytes by Staphylococcal Enterotoxin B Correlates with Toxin-Induced Proliferation and Is Regulated through Protein Kinase C

Journal: Infection and Immunity

doi:

FACScan analysis of TNF-α expression in monocytes and lymphocytes. Monocytes and lymphocytes (5 × 10 5 each) were mixed and incubated for 6 h with 100 ng of SEB per ml in the presence of 2 μM monensin. Samples were fixed and then permeabilized and stained with cell-type-specific antibody and analyzed on a FACScan flow cytometer. Negative controls (A and C) were without SEB. Samples were stained with anti-CD3 or anti-CD14 antisera to identify the following: lymphocytes (A), lymphocytes plus SEB (B), monocytes (C), and monocytes plus SEB (D).
Figure Legend Snippet: FACScan analysis of TNF-α expression in monocytes and lymphocytes. Monocytes and lymphocytes (5 × 10 5 each) were mixed and incubated for 6 h with 100 ng of SEB per ml in the presence of 2 μM monensin. Samples were fixed and then permeabilized and stained with cell-type-specific antibody and analyzed on a FACScan flow cytometer. Negative controls (A and C) were without SEB. Samples were stained with anti-CD3 or anti-CD14 antisera to identify the following: lymphocytes (A), lymphocytes plus SEB (B), monocytes (C), and monocytes plus SEB (D).

Techniques Used: Expressing, Incubation, Staining, Flow Cytometry, Cytometry

26) Product Images from "SPLIT TOLERANCE INDUCED BY ORTHOTOPIC LIVER TRANSPLANTATION IN MICE 1"

Article Title: SPLIT TOLERANCE INDUCED BY ORTHOTOPIC LIVER TRANSPLANTATION IN MICE 1

Journal: Transplantation

doi:

FACScan analysis of V β usage. The frequency of V β 3, V β 5, V β 8, and V β 11 TCR was examined in the T cells isolated from the spleens of naive B10.BR (donor), naive B10 (recipient), and recipients of B10.BR livers (30 days after transplant). Isotype-matched irrelevant mAb was used as a negative control.
Figure Legend Snippet: FACScan analysis of V β usage. The frequency of V β 3, V β 5, V β 8, and V β 11 TCR was examined in the T cells isolated from the spleens of naive B10.BR (donor), naive B10 (recipient), and recipients of B10.BR livers (30 days after transplant). Isotype-matched irrelevant mAb was used as a negative control.

Techniques Used: Isolation, Negative Control

27) Product Images from "Biological Effects of c-Mer Receptor Tyrosine Kinase in Hematopoietic Cells Depend on the Grb2 Binding Site in the Receptor and Activation of NF-?B"

Article Title: Biological Effects of c-Mer Receptor Tyrosine Kinase in Hematopoietic Cells Depend on the Grb2 Binding Site in the Receptor and Activation of NF-?B

Journal: Molecular and Cellular Biology

doi:

Increased receptor expression after selection of cells in the absence of IL-3. Cells expressing the indicated receptors were maintained for 14 days in medium with (+) IL-3 (control cells) (filled-in curve) or without (−) IL-3 (selected surviving cells) (unfilled curve). The expression of the receptors was detected by FACScan analysis using an FITC-conjugated anti-CD8 antibody. A scheme of the selection process is shown above the receptor expression histograms.
Figure Legend Snippet: Increased receptor expression after selection of cells in the absence of IL-3. Cells expressing the indicated receptors were maintained for 14 days in medium with (+) IL-3 (control cells) (filled-in curve) or without (−) IL-3 (selected surviving cells) (unfilled curve). The expression of the receptors was detected by FACScan analysis using an FITC-conjugated anti-CD8 antibody. A scheme of the selection process is shown above the receptor expression histograms.

Techniques Used: Expressing, Selection

28) Product Images from "Augmented efficacy of brentuximab vedotin combined with ruxolitinib and/or Navitoclax in a murine model of human Hodgkin’s lymphoma"

Article Title: Augmented efficacy of brentuximab vedotin combined with ruxolitinib and/or Navitoclax in a murine model of human Hodgkin’s lymphoma

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1524668113

The combination of ruxolitinib, Navitoclax, and BV triggers a mitochondrial apoptosis pathway in HDLM-2 cells. The mitochondrial membrane potential was measured by the membrane-permeant JC-1 dye staining. After treatment (ruxolitinib, 250 nM; Navitoclax, 50 nM; and BV, 100 ng/mL) for 48 h, the HDLM-2 cells were incubated with JC-1 at a concentration of 10 µg/mL in medium at 37 °C, 5% CO 2 for 10 min and then washed with PBS. The samples were collected on a FACScan flow cytometer. Data are representative of three independent experiments.
Figure Legend Snippet: The combination of ruxolitinib, Navitoclax, and BV triggers a mitochondrial apoptosis pathway in HDLM-2 cells. The mitochondrial membrane potential was measured by the membrane-permeant JC-1 dye staining. After treatment (ruxolitinib, 250 nM; Navitoclax, 50 nM; and BV, 100 ng/mL) for 48 h, the HDLM-2 cells were incubated with JC-1 at a concentration of 10 µg/mL in medium at 37 °C, 5% CO 2 for 10 min and then washed with PBS. The samples were collected on a FACScan flow cytometer. Data are representative of three independent experiments.

Techniques Used: Staining, Incubation, Concentration Assay, Flow Cytometry, Cytometry

29) Product Images from "Polydatin, a natural precursor of resveratrol, induces cell cycle arrest and differentiation of human colorectal Caco-2 cell"

Article Title: Polydatin, a natural precursor of resveratrol, induces cell cycle arrest and differentiation of human colorectal Caco-2 cell

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-264

Effect of ISBn alone or in combination on mithocondrial superoxide anions, nitric oxide, and enzyme scavengers activity in Caco-2 cells. (A) After 24 h Caco-2 treatment with polydatin and resveratrol alone or in combination (240 and 100 μM) at 37°C, the mitochondrial superoxide anion production was analysed by HE (20 ng · mL-1) staining. Dye accumulation was analysed by FACScan flow cytometer (FACScan, Becton Dickinson) by the CellQuest software, and the intensities of the bands were expressed as percent control. For each sample, 2 × 10 4 events were acquired. (B) Nitric oxide was measured in medium. Whole cell homogenates were used for evaluated the Catalase (C) and Mn-SOD activity (D) . The bars represent means ± SEM of three independent experiments. Asterisks indicate significant difference between the Caco-2-treated samples compared with control value ** P
Figure Legend Snippet: Effect of ISBn alone or in combination on mithocondrial superoxide anions, nitric oxide, and enzyme scavengers activity in Caco-2 cells. (A) After 24 h Caco-2 treatment with polydatin and resveratrol alone or in combination (240 and 100 μM) at 37°C, the mitochondrial superoxide anion production was analysed by HE (20 ng · mL-1) staining. Dye accumulation was analysed by FACScan flow cytometer (FACScan, Becton Dickinson) by the CellQuest software, and the intensities of the bands were expressed as percent control. For each sample, 2 × 10 4 events were acquired. (B) Nitric oxide was measured in medium. Whole cell homogenates were used for evaluated the Catalase (C) and Mn-SOD activity (D) . The bars represent means ± SEM of three independent experiments. Asterisks indicate significant difference between the Caco-2-treated samples compared with control value ** P

Techniques Used: Activity Assay, Staining, Flow Cytometry, Cytometry, Software

30) Product Images from "Valproic Acid Addresses Neuroendocrine Differentiation of LNCaP Cells and Maintains Cell Survival"

Article Title: Valproic Acid Addresses Neuroendocrine Differentiation of LNCaP Cells and Maintains Cell Survival

Journal: Drug Design, Development and Therapy

doi: 10.2147/DDDT.S229930

Effects of valproic acid on cell cycle distribution in prostate cancer cells. Cell cycle profile of LNCaP cells treated for 48, 72 and 96 hrs with vehicle (C) or 1mM of VPA ( A ). The latter condition at 96 hrs was reproduced in two other plates, drug-withdrawal and incubated for 72 and 96 hrs with refreshed medium (RM) ( C ), as described in the 'Material and Methods section. Cells were stained with propidium iodide and analyzed on a FACScan flow cytometer. Quantitative analysis of percentage gated cells at G0/G1, S and G2/M phases in the above reported experimental conditions ( B and D respectively). The results are representative of three independent experiments, with similar results.
Figure Legend Snippet: Effects of valproic acid on cell cycle distribution in prostate cancer cells. Cell cycle profile of LNCaP cells treated for 48, 72 and 96 hrs with vehicle (C) or 1mM of VPA ( A ). The latter condition at 96 hrs was reproduced in two other plates, drug-withdrawal and incubated for 72 and 96 hrs with refreshed medium (RM) ( C ), as described in the 'Material and Methods section. Cells were stained with propidium iodide and analyzed on a FACScan flow cytometer. Quantitative analysis of percentage gated cells at G0/G1, S and G2/M phases in the above reported experimental conditions ( B and D respectively). The results are representative of three independent experiments, with similar results.

Techniques Used: Incubation, Staining, Flow Cytometry, Cytometry

31) Product Images from "Induction of ASC pyroptosis requires gasdermin D or caspase-1/11-dependent mediators and IFNβ from pyroptotic macrophages"

Article Title: Induction of ASC pyroptosis requires gasdermin D or caspase-1/11-dependent mediators and IFNβ from pyroptotic macrophages

Journal: Cell Death & Disease

doi: 10.1038/s41419-020-2664-0

Pyroptosis reduces the antibacterial ability of ASCs. LPS-primed ASCs were incubated with SC3, SLN3, SCB, and SLNB respectively for 2 h, followed by infected with GFP-PAO1 at MOI of 1 for an additional 2 h. n = 3. a CFU counts were performed to determine the number of viable P. aeruginosa in the supernatants. b The intracellular GFP-fluorescence of ASCs was measured by a FACScan flow cytometer. The median fluorescence intensity represents the intracellular GFP-PAO1. n = 3. Statistical significance was determined using one-way analysis of variance. Data are shown as mean ± SD. ns: p > 0.05, * p
Figure Legend Snippet: Pyroptosis reduces the antibacterial ability of ASCs. LPS-primed ASCs were incubated with SC3, SLN3, SCB, and SLNB respectively for 2 h, followed by infected with GFP-PAO1 at MOI of 1 for an additional 2 h. n = 3. a CFU counts were performed to determine the number of viable P. aeruginosa in the supernatants. b The intracellular GFP-fluorescence of ASCs was measured by a FACScan flow cytometer. The median fluorescence intensity represents the intracellular GFP-PAO1. n = 3. Statistical significance was determined using one-way analysis of variance. Data are shown as mean ± SD. ns: p > 0.05, * p

Techniques Used: Incubation, Infection, Fluorescence, Flow Cytometry

32) Product Images from "Stable expression of constitutively-activated STAT3 in benign prostatic epithelial cells changes their phenotype to that resembling malignant cells"

Article Title: Stable expression of constitutively-activated STAT3 in benign prostatic epithelial cells changes their phenotype to that resembling malignant cells

Journal: Molecular Cancer

doi: 10.1186/1476-4598-4-2

Functional activity of STAT3 in S3c-transfected cells. Panel A: To show the functional activity of STAT3 expressed by the S3c gene, BPH-1 cells stably transfected with pIRES-S3c were treated with either 125 nM sense or antisense STAT3 oligonucleotide. Percent viability over time was determined by staining with propidium iodide, then quantifying fluorescence on a FACScan flow cytometer. Panel B: Treatment with 125 nM antisense STAT3 oligo reduced the amount of intracellular STAT3 protein in the clone of BPH-S3c cells shown in 3A. Twenty-four hours after transfection, BPH-S3c cells were harvested, fixed, and permeabilized, then stained with antibody to STAT3, as described in Materials and Methods. Quantification was performed on a FACScan flow cytometer. The black line indicates the amount of intracellular STAT3 in BPH-S3c cells treated with sense STAT3, while the grey line shows the amount of STAT3 in BPH-S3c cells given antisense STAT3. STAT3 expression was reduced by 66% in this experiment.
Figure Legend Snippet: Functional activity of STAT3 in S3c-transfected cells. Panel A: To show the functional activity of STAT3 expressed by the S3c gene, BPH-1 cells stably transfected with pIRES-S3c were treated with either 125 nM sense or antisense STAT3 oligonucleotide. Percent viability over time was determined by staining with propidium iodide, then quantifying fluorescence on a FACScan flow cytometer. Panel B: Treatment with 125 nM antisense STAT3 oligo reduced the amount of intracellular STAT3 protein in the clone of BPH-S3c cells shown in 3A. Twenty-four hours after transfection, BPH-S3c cells were harvested, fixed, and permeabilized, then stained with antibody to STAT3, as described in Materials and Methods. Quantification was performed on a FACScan flow cytometer. The black line indicates the amount of intracellular STAT3 in BPH-S3c cells treated with sense STAT3, while the grey line shows the amount of STAT3 in BPH-S3c cells given antisense STAT3. STAT3 expression was reduced by 66% in this experiment.

Techniques Used: Functional Assay, Activity Assay, Transfection, Stable Transfection, Staining, Fluorescence, Flow Cytometry, Cytometry, Expressing

33) Product Images from "p62 Suppressed VK3-induced Oxidative Damage Through Keap1/Nrf2 Pathway In Human Ovarian Cancer Cells"

Article Title: p62 Suppressed VK3-induced Oxidative Damage Through Keap1/Nrf2 Pathway In Human Ovarian Cancer Cells

Journal: Journal of Cancer

doi: 10.7150/jca.34423

VK3 promoted the apoptosis of SKOV3 ovarian cancer cells. (A) SKOV3 and SKOV3/DDP cells were treated with VK3 for 8 and 16 h. The MTT assay was used to examine the cell viability. Data are presented as mean ± SD, n = 3. (B) Both cells were treated with 15 µM VK3 and stained with Hoechst 33342. Cell morphology was observed by fluorescence microscopy. Arrows indicate apoptotic cells. Scale bar = 20 µm. (C) Cells were stained with Annexin V-FITC/PI, and the ratio of apoptosis was detected using a FACScan flow cytometer. Data are presented as mean ± SD, n = 3. (D) Apoptotic rate in (C) was quantified in both cells. Data are presented as mean ± SD, n = 3. * P
Figure Legend Snippet: VK3 promoted the apoptosis of SKOV3 ovarian cancer cells. (A) SKOV3 and SKOV3/DDP cells were treated with VK3 for 8 and 16 h. The MTT assay was used to examine the cell viability. Data are presented as mean ± SD, n = 3. (B) Both cells were treated with 15 µM VK3 and stained with Hoechst 33342. Cell morphology was observed by fluorescence microscopy. Arrows indicate apoptotic cells. Scale bar = 20 µm. (C) Cells were stained with Annexin V-FITC/PI, and the ratio of apoptosis was detected using a FACScan flow cytometer. Data are presented as mean ± SD, n = 3. (D) Apoptotic rate in (C) was quantified in both cells. Data are presented as mean ± SD, n = 3. * P

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

34) Product Images from "Aciculatin Inhibits Granulocyte Colony-Stimulating Factor Production by Human Interleukin 1?-Stimulated Fibroblast-Like Synoviocytes"

Article Title: Aciculatin Inhibits Granulocyte Colony-Stimulating Factor Production by Human Interleukin 1?-Stimulated Fibroblast-Like Synoviocytes

Journal: PLoS ONE

doi: 10.1371/journal.pone.0042389

Aciculatin inhibits the phosphorylation of JAK, STAT, and Akt in FLS and the differentiation of 32Dcl3 cells. (A, C, and D) FLS were incubated with 0, 1, 3, or 10 µM aciculatin (A1, A3, and A10) for 30 min, and then for 24 h with 10 ng/mL of IL-1β in the continued presence of aciculatin. Whole cell extracts were then prepared for western blot analysis for the indicated proteins (A and C); equal amounts of cell culture media (“conditioned medium”) were collected and concentrated 10-fold (v/v) (lanes 1–4) or PBS only (lane 5), and then immunoprecipitated with 1 µg of anti-G-CSF antibody, followed by immunoblot analysis using anti-G-CSF antibody or anti-β-actin antibody (as an internal control) (D). (B) FLS were incubated with 0 or 10 µM aciculatin for 30 min, and then for 1 h with 10 ng/mL of IL-1β in the continued presence of aciculatin. The DNA binding activity of the nuclear extracts was then examined in an electrophoretic mobility shift assay using a specific STAT3 DNA probe. (E) Ten-fold concentrated conditioned medium was prepared from FLS incubated with or without aciculatin, and then with IL-1β as in (D), or with IL-1β plus an anti-G-CSF antibody. 32Dcl3 cells were incubated for 10 days with a medium containing 50% of these different conditioned mediums. The cells were then were subjected to Wright-Giemsa staining to detect neutrophils (top row) or washed twice with PBS, incubated at 4°C for 45 min with anti-CD11b FITC-conjugated and anti-CD11a/CD18 PE-conjugated antibodies, and their fluorescence was analyzed by FACScan flow cytometry (bottom row). Magnification = ×100; scale bar = 20 µm. In (A) and (C), the extents of indicated proteins expression were quantitated using a densitometer with the Image-Pro plus software, and the relative levels were calculated as the ratios of proteins to GAPDH or β-actin protein levels. The results are expressed as the mean ± SEM, with n = 3. * p
Figure Legend Snippet: Aciculatin inhibits the phosphorylation of JAK, STAT, and Akt in FLS and the differentiation of 32Dcl3 cells. (A, C, and D) FLS were incubated with 0, 1, 3, or 10 µM aciculatin (A1, A3, and A10) for 30 min, and then for 24 h with 10 ng/mL of IL-1β in the continued presence of aciculatin. Whole cell extracts were then prepared for western blot analysis for the indicated proteins (A and C); equal amounts of cell culture media (“conditioned medium”) were collected and concentrated 10-fold (v/v) (lanes 1–4) or PBS only (lane 5), and then immunoprecipitated with 1 µg of anti-G-CSF antibody, followed by immunoblot analysis using anti-G-CSF antibody or anti-β-actin antibody (as an internal control) (D). (B) FLS were incubated with 0 or 10 µM aciculatin for 30 min, and then for 1 h with 10 ng/mL of IL-1β in the continued presence of aciculatin. The DNA binding activity of the nuclear extracts was then examined in an electrophoretic mobility shift assay using a specific STAT3 DNA probe. (E) Ten-fold concentrated conditioned medium was prepared from FLS incubated with or without aciculatin, and then with IL-1β as in (D), or with IL-1β plus an anti-G-CSF antibody. 32Dcl3 cells were incubated for 10 days with a medium containing 50% of these different conditioned mediums. The cells were then were subjected to Wright-Giemsa staining to detect neutrophils (top row) or washed twice with PBS, incubated at 4°C for 45 min with anti-CD11b FITC-conjugated and anti-CD11a/CD18 PE-conjugated antibodies, and their fluorescence was analyzed by FACScan flow cytometry (bottom row). Magnification = ×100; scale bar = 20 µm. In (A) and (C), the extents of indicated proteins expression were quantitated using a densitometer with the Image-Pro plus software, and the relative levels were calculated as the ratios of proteins to GAPDH or β-actin protein levels. The results are expressed as the mean ± SEM, with n = 3. * p

Techniques Used: Incubation, Western Blot, Cell Culture, Immunoprecipitation, Binding Assay, Activity Assay, Electrophoretic Mobility Shift Assay, Staining, Fluorescence, Flow Cytometry, Cytometry, Expressing, Software

35) Product Images from "Shigella flexneri IpaH7.8 Facilitates Escape of Virulent Bacteria from the Endocytic Vacuoles of Mouse and Human Macrophages"

Article Title: Shigella flexneri IpaH7.8 Facilitates Escape of Virulent Bacteria from the Endocytic Vacuoles of Mouse and Human Macrophages

Journal: Infection and Immunity

doi:

DNA analysis by flow cytometry. J774 cells incubated with Shigella strains were lysed, and the released nuclei were labeled with propidium iodide and analyzed on a FACScan flow cytometer. Cells were infected with 2457T (A), M4243A1 (B), or pWR700 (C) or were noninfected (D).
Figure Legend Snippet: DNA analysis by flow cytometry. J774 cells incubated with Shigella strains were lysed, and the released nuclei were labeled with propidium iodide and analyzed on a FACScan flow cytometer. Cells were infected with 2457T (A), M4243A1 (B), or pWR700 (C) or were noninfected (D).

Techniques Used: Flow Cytometry, Cytometry, Incubation, Labeling, Infection

36) Product Images from "Vacuolating Cytotoxin of Helicobacter pylori Induces Apoptosis in the Human Gastric Epithelial Cell Line AGS"

Article Title: Vacuolating Cytotoxin of Helicobacter pylori Induces Apoptosis in the Human Gastric Epithelial Cell Line AGS

Journal: Infection and Immunity

doi: 10.1128/IAI.69.8.5080-5087.2001

(a) Western blot analysis of VacA. Fifty-microliter samples of concentrated supernatant from H. pylori P12 and P14 were subjected to SDS-PAGE and immunoblotted with anti-VacA antibodies. Native VacA appeared (at approximately 87 kDa) only with the supernatant of strain P12. (b) FACScan analysis of AGS cells. AGS cells were treated for 48 h with 250 μl of supernatant of H. pylori strains P12 and P14/ml. The abscissa displays the counts and the ordinate displays the fluorescence on a logarithmic scale. Apoptotic cells appear as a sub-G 1 peak measured through the marker region (M1). Treatment with concentrated supernatant of the vacA wild-type strain P12 increased the amount of apoptotic cells compared to untreated AGS cells or AGS cells treated with the isogenic vacA mutant strain P14.
Figure Legend Snippet: (a) Western blot analysis of VacA. Fifty-microliter samples of concentrated supernatant from H. pylori P12 and P14 were subjected to SDS-PAGE and immunoblotted with anti-VacA antibodies. Native VacA appeared (at approximately 87 kDa) only with the supernatant of strain P12. (b) FACScan analysis of AGS cells. AGS cells were treated for 48 h with 250 μl of supernatant of H. pylori strains P12 and P14/ml. The abscissa displays the counts and the ordinate displays the fluorescence on a logarithmic scale. Apoptotic cells appear as a sub-G 1 peak measured through the marker region (M1). Treatment with concentrated supernatant of the vacA wild-type strain P12 increased the amount of apoptotic cells compared to untreated AGS cells or AGS cells treated with the isogenic vacA mutant strain P14.

Techniques Used: Western Blot, SDS Page, Fluorescence, Marker, Mutagenesis

FACScan analysis of AGS cells. AGS cells were incubated for 60 h with 100 μg of recombinant VacA/ml. The abscissa displays the counts, and the ordinate displays the fluorescence on a logarithmic scale. Apoptotic cells appear as a sub-G 1 peak, which was statistically acquired through the marker region (M1). Incubation with recombinant VacA increases apoptotic cell death (right).
Figure Legend Snippet: FACScan analysis of AGS cells. AGS cells were incubated for 60 h with 100 μg of recombinant VacA/ml. The abscissa displays the counts, and the ordinate displays the fluorescence on a logarithmic scale. Apoptotic cells appear as a sub-G 1 peak, which was statistically acquired through the marker region (M1). Incubation with recombinant VacA increases apoptotic cell death (right).

Techniques Used: Incubation, Recombinant, Fluorescence, Marker

37) Product Images from "Death Receptor-Induced Activation of the Chk2- and Histone H2AX-Associated DNA Damage Response Pathways ▿"

Article Title: Death Receptor-Induced Activation of the Chk2- and Histone H2AX-Associated DNA Damage Response Pathways ▿

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.00581-08

H2AX is phosphorylated by DNA-PK and Chk2 is phosphorylated by both DNA-PK and ATM in response to TRAIL. (A) Effect of the DNA-PKi NU7441 on the phosphorylations of Chk2 and H2AX after TRAIL treatment. HCT116 cells were treated with the DNA-PKi (1 h) prior to the addition of TRAIL. Protein phosphorylations (P-Chk2 T68 and γ-H2AX) were analyzed by Western blotting. Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated. (B) Effect of the ATMi KU-55933 on the phosphorylations of Chk2 and H2AX after TRAIL treatment. HCT116 cells were treated with the ATMi (1 h) prior to the addition of TRAIL. Protein phosphorylations (P-Chk2 T68 and γ-H2AX) were analyzed by Western blotting. Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated. (C) Inhibition of γ-H2AX formation by the DNA-PKi (10 μM, 1 h) in TRAIL-treated HCT116 cells (0.1 μg/ml, 3 h). Cells were analyzed by FACScan flow cytometry. The x axis indicates γ-H2AX content (γ-H2AX-positive cells are circled in gray), and the y axis indicates the number of cells. The percentage of γ-H2AX-positive cells was significantly reduced by pretreatment with the DNA-PKi ( n = 3; unpaired t test, P
Figure Legend Snippet: H2AX is phosphorylated by DNA-PK and Chk2 is phosphorylated by both DNA-PK and ATM in response to TRAIL. (A) Effect of the DNA-PKi NU7441 on the phosphorylations of Chk2 and H2AX after TRAIL treatment. HCT116 cells were treated with the DNA-PKi (1 h) prior to the addition of TRAIL. Protein phosphorylations (P-Chk2 T68 and γ-H2AX) were analyzed by Western blotting. Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated. (B) Effect of the ATMi KU-55933 on the phosphorylations of Chk2 and H2AX after TRAIL treatment. HCT116 cells were treated with the ATMi (1 h) prior to the addition of TRAIL. Protein phosphorylations (P-Chk2 T68 and γ-H2AX) were analyzed by Western blotting. Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated. (C) Inhibition of γ-H2AX formation by the DNA-PKi (10 μM, 1 h) in TRAIL-treated HCT116 cells (0.1 μg/ml, 3 h). Cells were analyzed by FACScan flow cytometry. The x axis indicates γ-H2AX content (γ-H2AX-positive cells are circled in gray), and the y axis indicates the number of cells. The percentage of γ-H2AX-positive cells was significantly reduced by pretreatment with the DNA-PKi ( n = 3; unpaired t test, P

Techniques Used: Western Blot, Staining, Inhibition, Flow Cytometry, Cytometry

Effect of TRAIL on DDR proteins is observed at all phases of the cell cycle and is not restricted to HCT116 cells. (A) Relationships between γ-H2AX cellular content and cell cycle. Untreated (upper left panel) and TRAIL-treated (0.1 μg/ml, 2 h) HCT116 cells (upper right panel) were analyzed by FACScan flow cytometry. The x axis indicates the DNA content (as determined by propidium iodide staining), and the y axis indicates γ-H2AX content (cells positive for γ-H2AX are in red, and the negative cells are in gray). Results are quantitated in the lower panel. (B) Effects of TRAIL on Chk2, ATM, H2AX, and DNA-PK phosphorylations in HeLa cells. Protein phosphorylations were analyzed by Western blotting. The asterisk corresponds to an unspecific cross-reactive protein for the phospho-T68-Chk2 antibody (P-Chk2 T68). Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated at the bottom. (C) Activation of Chk2 in response to TRAIL in Jurkat cells. Cells were treated with 0.1 μg/ml TRAIL for the indicated times, and the phosphorylation of Chk2 on T68 was analyzed by Western blotting. Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated.
Figure Legend Snippet: Effect of TRAIL on DDR proteins is observed at all phases of the cell cycle and is not restricted to HCT116 cells. (A) Relationships between γ-H2AX cellular content and cell cycle. Untreated (upper left panel) and TRAIL-treated (0.1 μg/ml, 2 h) HCT116 cells (upper right panel) were analyzed by FACScan flow cytometry. The x axis indicates the DNA content (as determined by propidium iodide staining), and the y axis indicates γ-H2AX content (cells positive for γ-H2AX are in red, and the negative cells are in gray). Results are quantitated in the lower panel. (B) Effects of TRAIL on Chk2, ATM, H2AX, and DNA-PK phosphorylations in HeLa cells. Protein phosphorylations were analyzed by Western blotting. The asterisk corresponds to an unspecific cross-reactive protein for the phospho-T68-Chk2 antibody (P-Chk2 T68). Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated at the bottom. (C) Activation of Chk2 in response to TRAIL in Jurkat cells. Cells were treated with 0.1 μg/ml TRAIL for the indicated times, and the phosphorylation of Chk2 on T68 was analyzed by Western blotting. Tubulin was used as a loading control. The percentage of apoptosis measured by Hoechst staining is indicated.

Techniques Used: Flow Cytometry, Cytometry, Staining, Western Blot, Activation Assay

38) Product Images from "Saccharomyces boulardii Improves Intestinal Epithelial Cell Restitution by Inhibiting ?v?5 Integrin Activation State"

Article Title: Saccharomyces boulardii Improves Intestinal Epithelial Cell Restitution by Inhibiting ?v?5 Integrin Activation State

Journal: PLoS ONE

doi: 10.1371/journal.pone.0045047

Impact of Sb S on integrin expression. Sub-confluent HCT-8/E11 cells were incubated for 5 h with (dark bars) or without (light bars) Sb S, then harvested and resuspended in the presence of anti-integrin subunit mAbs. After incubation with the appropriate secondary Alexa 488-conjugated Ab, cell-bound fluorescence was quantified using a Becton-Dickinson FACScan flow cytometer. Non-specific labeling was determined by incubating cells with the secondary Alexa 488-conjugated Ab alone (Ctrl). Data represent the mean+SD of 6 separate experiments.
Figure Legend Snippet: Impact of Sb S on integrin expression. Sub-confluent HCT-8/E11 cells were incubated for 5 h with (dark bars) or without (light bars) Sb S, then harvested and resuspended in the presence of anti-integrin subunit mAbs. After incubation with the appropriate secondary Alexa 488-conjugated Ab, cell-bound fluorescence was quantified using a Becton-Dickinson FACScan flow cytometer. Non-specific labeling was determined by incubating cells with the secondary Alexa 488-conjugated Ab alone (Ctrl). Data represent the mean+SD of 6 separate experiments.

Techniques Used: Expressing, Incubation, Fluorescence, Flow Cytometry, Cytometry, Labeling

39) Product Images from "The brown adipocyte differentiation pathway in birds: An evolutionary road not taken"

Article Title: The brown adipocyte differentiation pathway in birds: An evolutionary road not taken

Journal: BMC Biology

doi: 10.1186/1741-7007-6-17

ABALC cellular phenotype is associated with increased mitochondrial volume . FACScan of cells stained with JC-1. FL2-H is red fluorescence. Black line, unstained mesenchymal cells; blue line, stained mesenchymal cells; red line, limb bud preadipocytes (LBPAs); pink line, limb bud mesenchyme (LBM) adipocytes grown in chicken serum; green line, avian brown adipocyte-like cells (ABALCs).
Figure Legend Snippet: ABALC cellular phenotype is associated with increased mitochondrial volume . FACScan of cells stained with JC-1. FL2-H is red fluorescence. Black line, unstained mesenchymal cells; blue line, stained mesenchymal cells; red line, limb bud preadipocytes (LBPAs); pink line, limb bud mesenchyme (LBM) adipocytes grown in chicken serum; green line, avian brown adipocyte-like cells (ABALCs).

Techniques Used: Staining, Fluorescence

40) Product Images from "A Single Immunoglobulin-like Domain of the Human Neural Cell Adhesion Molecule L1 Supports Adhesion by Multiple Vascular and Platelet Integrins "

Article Title: A Single Immunoglobulin-like Domain of the Human Neural Cell Adhesion Molecule L1 Supports Adhesion by Multiple Vascular and Platelet Integrins

Journal: The Journal of Cell Biology

doi:

( A ) Integrin profiles of wild-type ECV304 human umbilical vein endothelial cells or ECV304 cells repeatedly sorted for a lack of α v β 3 expression. ( B and C ) α v β 3 - and α v β 1 -dependent adhesion of wild-type or sorted ECV304 endothelial cells to L1-Ig6. ( A ) Integrin expression is represented by FACS ® histograms. Cells were treated with antibodies to α v integrins (mAb 17E6), to α v β 3 (mAb LM609), to α v or β 3 integrin subunits (polyclonal VNR), or to β 1 integrins (mAb P4C10). These cells were subsequently stained with fluorescein-conjugated goat anti–mouse or goat anti– rabbit antibodies and were analyzed using a FACScan ® flow cytometer. Control cells were treated with secondary fluorescein-conjugated antibody only. ( B and C ) Wild-type or sorted cells were allowed to adhere to immobilized L1-Ig6 fusion protein offered at 40 μg/ml. Adhesion was performed in the presence of Ca 2+ (2 mM), Mg 2+ (2 mM), and Mn 2+ (0.4 mM). Some cells were pretreated with antibody to α v β 3 (mAb LM609; 80 μg/ml), to β 1 integrins (mAb P4C10; 80 μg/ml), to α v integrins (MAb 17E6; 80 μg/ml), or with a combination of antibodies. After 40 min non- adherent cells were removed by washing and the remaining adherent cells counted per unit area with a ×15 high powered objective. Both , a combination of the two preceding antibodies. Experimental treatments were performed in triplicate with a minimum of four areas counted per well. Error bars represent ±1 SD.
Figure Legend Snippet: ( A ) Integrin profiles of wild-type ECV304 human umbilical vein endothelial cells or ECV304 cells repeatedly sorted for a lack of α v β 3 expression. ( B and C ) α v β 3 - and α v β 1 -dependent adhesion of wild-type or sorted ECV304 endothelial cells to L1-Ig6. ( A ) Integrin expression is represented by FACS ® histograms. Cells were treated with antibodies to α v integrins (mAb 17E6), to α v β 3 (mAb LM609), to α v or β 3 integrin subunits (polyclonal VNR), or to β 1 integrins (mAb P4C10). These cells were subsequently stained with fluorescein-conjugated goat anti–mouse or goat anti– rabbit antibodies and were analyzed using a FACScan ® flow cytometer. Control cells were treated with secondary fluorescein-conjugated antibody only. ( B and C ) Wild-type or sorted cells were allowed to adhere to immobilized L1-Ig6 fusion protein offered at 40 μg/ml. Adhesion was performed in the presence of Ca 2+ (2 mM), Mg 2+ (2 mM), and Mn 2+ (0.4 mM). Some cells were pretreated with antibody to α v β 3 (mAb LM609; 80 μg/ml), to β 1 integrins (mAb P4C10; 80 μg/ml), to α v integrins (MAb 17E6; 80 μg/ml), or with a combination of antibodies. After 40 min non- adherent cells were removed by washing and the remaining adherent cells counted per unit area with a ×15 high powered objective. Both , a combination of the two preceding antibodies. Experimental treatments were performed in triplicate with a minimum of four areas counted per well. Error bars represent ±1 SD.

Techniques Used: Expressing, FACS, Staining, Flow Cytometry, Cytometry

( A ) Integrin profiles of wild-type CHO-K1 cells, of CHO cells transfected with α IIb and β 3 subunits ( A5 ) and of CHO cells transfected to express active α IIb β 3 (α IIb α L Δβ 3 cells). ( B ) Concentration-dependent adhesion of CHO-K1, A5, and α IIb α L Δβ 3 cells to L1-Ig6. ( A ) Integrin expression is represented by FACS ® histograms. Cells were treated with antibodies to hamster α 5 β 1 (monoclonal PB1), to human α v β 3 (mAb LM609), to human α IIb β 3 (mAb LJ-CP8), to hamster or human α v or β 3 integrin subunits (polyclonal VNR), or to human β 3 integrins (mAb 7E3). These cells were subsequently stained with fluorescein-conjugated goat anti–mouse or goat anti–rabbit antibodies and were analyzed using a FACScan ® flow cytometer. Control cells were treated with secondary fluorescein-conjugated antibody only. ( B ) Wild-type or transfected cells were allowed to adhere to immobilized L1-Ig6 fusion protein offered at concentrations ranging from 1 to 100 μg/ml. After 30 min non-adherent cells were removed by washing and the remaining adherent cells counted per unit area with a ×15 high powered objective. Experimental treatments were performed in triplicate with a minimum of four areas counted per well. No significant adhesion was observed on GST alone and any minimal residual adhesion to BSA-blocked plastic alone has been subtracted from the cell counts shown. Error bars represent ±1 SD.
Figure Legend Snippet: ( A ) Integrin profiles of wild-type CHO-K1 cells, of CHO cells transfected with α IIb and β 3 subunits ( A5 ) and of CHO cells transfected to express active α IIb β 3 (α IIb α L Δβ 3 cells). ( B ) Concentration-dependent adhesion of CHO-K1, A5, and α IIb α L Δβ 3 cells to L1-Ig6. ( A ) Integrin expression is represented by FACS ® histograms. Cells were treated with antibodies to hamster α 5 β 1 (monoclonal PB1), to human α v β 3 (mAb LM609), to human α IIb β 3 (mAb LJ-CP8), to hamster or human α v or β 3 integrin subunits (polyclonal VNR), or to human β 3 integrins (mAb 7E3). These cells were subsequently stained with fluorescein-conjugated goat anti–mouse or goat anti–rabbit antibodies and were analyzed using a FACScan ® flow cytometer. Control cells were treated with secondary fluorescein-conjugated antibody only. ( B ) Wild-type or transfected cells were allowed to adhere to immobilized L1-Ig6 fusion protein offered at concentrations ranging from 1 to 100 μg/ml. After 30 min non-adherent cells were removed by washing and the remaining adherent cells counted per unit area with a ×15 high powered objective. Experimental treatments were performed in triplicate with a minimum of four areas counted per well. No significant adhesion was observed on GST alone and any minimal residual adhesion to BSA-blocked plastic alone has been subtracted from the cell counts shown. Error bars represent ±1 SD.

Techniques Used: Transfection, Concentration Assay, Expressing, FACS, Staining, Flow Cytometry, Cytometry

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Flow Cytometry:

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Article Title: A Cyclooxygenase-2 Inhibitor (SC-58125) Blocks Growth of Established Human Colon Cancer Xenografts 1
Article Snippet: .. Cells were then filtered through 50- µ m mesh immediately prior to analysis on a Becton Dickinson FACScan flow cytometer. ..

Article Title: MD41, a novel T helper 0 clone, mediates mast-cell dependent delayed-type hypersensitivity in mice
Article Snippet: .. Finally, after another two washes with HBSS/NaN3 , the cells were analyzed using a FACScan flow cytometer (Becton Dickinson, San Jose, CA). .. For the double staining of intracellular cytokines, MD41 cells were resuspended at 105 −106 /ml in Iscove's modified medium supplemented with 10% FCS and stimulated with con A at 5 µg/ml or phorbol 12-myristate 13-acetate (PMA) at 10 ng/ml for 48 hr.

Fluorescence:

Article Title: A Precise Nanostructure of Folate-Overhung Mitoxantrone DNA Tetrahedron for Targeted Capture Leukemia
Article Snippet: .. The fluorescence intensity was measured by the FACScan flow cytometer (Becton Dickinson Biosciences, Franklin Lakes, NJ, USA). ..

Cytometry:

Article Title: Antimonial-Mediated DNA Fragmentation in Leishmania infantum Amastigotes
Article Snippet: .. Cells were immediately analyzed on the FACScan flow cytometer (Becton Dickinson) using an argon-ion laser tuned to 488 nm. .. Green cell fluorescence, gated on forward and side-light scatter, was collected using a band-pass filter (525 ± 10 nm) and displayed using a logarithmic amplification (FL-1).

Article Title: Sustained Elevated Levels of VCAM-1 in Cultured Fibroblast-like Synoviocytes Can Be Achieved by TNF-? in Combination with Either IL-4 or IL-13 through Increased mRNA Stability
Article Snippet: .. Flow cytometry was performed using a FACScan flow cytometer (Becton Dickinson, Cowley, UK). ..

Article Title: Mechanisms Involved in Stimulation of Human Immunodeficiency Virus Type 1 Replication by Aminooxypentane RANTES
Article Snippet: .. After the final wash, cells were fixed with 300 μl of 1% paraformaldehyde and analyzed using a FacScan flow cytometer and Lysis II software (Becton Dickinson). .. The long terminal repeat (LTR) region of the HIV-1 D-92UG021 strain was PCR amplified from DNA of infected PBMC using LTR1 and LTR2 as external primers and S2-LTR4 and LTR3-LTR4 as nested primer pairs ( ).

Article Title: High-Throughput Multiplex Flow Cytometry Screening for Botulinum Neurotoxin Type A Light Chain Protease Inhibitors
Article Snippet: .. Samples were run on a Becton Dickinson FACScan flow cytometer with 100 µL samples run on the cytometer every 30 min. FACScan PMT settings were FSC E00/9.25, SSC 225 log, FL1 700 log and FL2 600 log, compensation values of FL2–12% FL1, and gates were drawn on individual bead set in the FL2 histogram plot. .. Percent inhibition was calculated by taking the 1 h sample value—1 h no inhibitor value and dividing by the 1 h no protease value—1 h no inhibitor value and multiplying by 100.

Article Title: Apoptosis and apoptosis-associated perturbations of peripheral blood lymphocytes during HIV infection: comparison between AIDS patients and asymptomatic long-term non-progressors
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Article Title: A Cyclooxygenase-2 Inhibitor (SC-58125) Blocks Growth of Established Human Colon Cancer Xenografts 1
Article Snippet: .. Cells were then filtered through 50- µ m mesh immediately prior to analysis on a Becton Dickinson FACScan flow cytometer. ..

Article Title: MD41, a novel T helper 0 clone, mediates mast-cell dependent delayed-type hypersensitivity in mice
Article Snippet: .. Finally, after another two washes with HBSS/NaN3 , the cells were analyzed using a FACScan flow cytometer (Becton Dickinson, San Jose, CA). .. For the double staining of intracellular cytokines, MD41 cells were resuspended at 105 −106 /ml in Iscove's modified medium supplemented with 10% FCS and stimulated with con A at 5 µg/ml or phorbol 12-myristate 13-acetate (PMA) at 10 ng/ml for 48 hr.

Incubation:

Article Title: Apoptosis and apoptosis-associated perturbations of peripheral blood lymphocytes during HIV infection: comparison between AIDS patients and asymptomatic long-term non-progressors
Article Snippet: .. Finally, the double-stained cells were incubated overnight at 4°C in the dark and were then analysed in their staining solution by a FACScan flow cytometer (Becton Dickinson). .. The green fluorescence was collected after a 530/30 BP nm filter, the red fluorescence from 7-AAD was filtered through a 650 long pass filter.

Lysis:

Article Title: Mechanisms Involved in Stimulation of Human Immunodeficiency Virus Type 1 Replication by Aminooxypentane RANTES
Article Snippet: .. After the final wash, cells were fixed with 300 μl of 1% paraformaldehyde and analyzed using a FacScan flow cytometer and Lysis II software (Becton Dickinson). .. The long terminal repeat (LTR) region of the HIV-1 D-92UG021 strain was PCR amplified from DNA of infected PBMC using LTR1 and LTR2 as external primers and S2-LTR4 and LTR3-LTR4 as nested primer pairs ( ).

Staining:

Article Title: Apoptosis and apoptosis-associated perturbations of peripheral blood lymphocytes during HIV infection: comparison between AIDS patients and asymptomatic long-term non-progressors
Article Snippet: .. Finally, the double-stained cells were incubated overnight at 4°C in the dark and were then analysed in their staining solution by a FACScan flow cytometer (Becton Dickinson). .. The green fluorescence was collected after a 530/30 BP nm filter, the red fluorescence from 7-AAD was filtered through a 650 long pass filter.

Software:

Article Title: Mechanisms Involved in Stimulation of Human Immunodeficiency Virus Type 1 Replication by Aminooxypentane RANTES
Article Snippet: .. After the final wash, cells were fixed with 300 μl of 1% paraformaldehyde and analyzed using a FacScan flow cytometer and Lysis II software (Becton Dickinson). .. The long terminal repeat (LTR) region of the HIV-1 D-92UG021 strain was PCR amplified from DNA of infected PBMC using LTR1 and LTR2 as external primers and S2-LTR4 and LTR3-LTR4 as nested primer pairs ( ).

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  • 94
    Becton Dickinson facscan flow cytometer
    SC-58125 does not induce apoptosis. LLC (A) or HCA-7 (B) cells untreated (negative control), treated with SC-58635 (positive control), DMSO (vehicle), or with 100 µM SC-58125 for 24 hours (LLC), or 72 hours (HCA-7). Cells were harvested then fixed, and fragmented DNA was labeled using the TUNEL assay with dUTP- FITC as substrate. Analysis was performed using a Becton Dickinson <t>FACScan</t> flow cytometer. Ten thousand gated events were analyzed.
    Facscan Flow Cytometer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 512 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson facsan flow cytometer
    Folate deficiency provokes apoptotic lethality in synoviocytes. HIG-82 synoviocytes (1.5×10 5 ) were plated in 60-mm cultured dishes for 24 h. The culture medium was replaced with FC, MFD, and FD media and then continued cultivating for additional 48 h. (A) Cells were then collected, washed with <t>PBS,</t> fixed in PBS-methanol (1:2 v/v) solution and maintained at 4°C for at least 18 h. After one washed with PBS, the cell pellets were then stained with a PI solution containing PBS, PI (40μg/mL), and DNase-free RNase A (40μg/mL) for 30 min at RT in the dark. The cell pellets were then analyzed using a Becton-Dickinson <t>FACSan</t> flowcytometer. The epirubicin (500 nM) treatment (Epi) is a positive control assay of apoptosis. The blank bar, gray bar, right slash bar and left slash bar represent FC, MFD, FD and Epi treatment, respectively. The percentages of subG1 population determined by the PI fluorescent intensity in apoptosis cells which was weaker than that of cells in the G1 phase. The percentages of apoptosis cells were characterized as the percentages of cells in the SubG1 region of the DNA distribution histograms. The FD subG1 bar graph is compared with FC or MFD. A p
    Facsan Flow Cytometer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 89/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson two color flow cytometry
    DPV022 inhibits Bax-induced apoptosis and Bax oligomerization. (A) HeLa cells were cotransfected with pEGFP, pEGFP-DPV022, pEGFP-FPV039, pEGFP-M11L, or pEGFP-Bcl-2 and pcDNA3-HA-Bax. Apoptosis was assessed by quantifying TMRE fluorescence via flow <t>cytometry,</t>
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    SC-58125 does not induce apoptosis. LLC (A) or HCA-7 (B) cells untreated (negative control), treated with SC-58635 (positive control), DMSO (vehicle), or with 100 µM SC-58125 for 24 hours (LLC), or 72 hours (HCA-7). Cells were harvested then fixed, and fragmented DNA was labeled using the TUNEL assay with dUTP- FITC as substrate. Analysis was performed using a Becton Dickinson FACScan flow cytometer. Ten thousand gated events were analyzed.

    Journal: Neoplasia (New York, N.Y.)

    Article Title: A Cyclooxygenase-2 Inhibitor (SC-58125) Blocks Growth of Established Human Colon Cancer Xenografts 1

    doi:

    Figure Lengend Snippet: SC-58125 does not induce apoptosis. LLC (A) or HCA-7 (B) cells untreated (negative control), treated with SC-58635 (positive control), DMSO (vehicle), or with 100 µM SC-58125 for 24 hours (LLC), or 72 hours (HCA-7). Cells were harvested then fixed, and fragmented DNA was labeled using the TUNEL assay with dUTP- FITC as substrate. Analysis was performed using a Becton Dickinson FACScan flow cytometer. Ten thousand gated events were analyzed.

    Article Snippet: Cells were then filtered through 50- µ m mesh immediately prior to analysis on a Becton Dickinson FACScan flow cytometer.

    Techniques: High Content Screening, Negative Control, Positive Control, Labeling, TUNEL Assay, Flow Cytometry, Cytometry

    Induced apoptosis in leukemia BALL-1 cells by folate-overhung mitoxantrone DNA tetrahedra. ( A–D ): Induced apoptosis after treatment with varying formulations at 10 h, and the cells were stained using Annexin V-Fluor 647 and 7AAD, then assessed according to manufacturer instructions by the FACScan flow cytometer. UR: Late apoptosis and dead cells; LR: Early apoptosis cells ( A ). PBS; ( B ). Free mitoxantrone; ( C ). mitoxantrone DNA tetrahedra; ( D ). Folate-overhung mitoxantrone DNA tetrahedra; ( E ). The early apoptosis rate in BALL-1 cells ( F ). The late apoptosis rate in BALL-1 cells. ( E–F ): 1. PBS; 2. Free mitoxantrone; 3. Mitoxantrone DNA tetrahedra; 4. Folate-overhung mitoxantrone DNA tetrahedra. a. vs. PBS; b. vs. mitoxantrone DNA tetrahedra, p

    Journal: Nanomaterials

    Article Title: A Precise Nanostructure of Folate-Overhung Mitoxantrone DNA Tetrahedron for Targeted Capture Leukemia

    doi: 10.3390/nano10050951

    Figure Lengend Snippet: Induced apoptosis in leukemia BALL-1 cells by folate-overhung mitoxantrone DNA tetrahedra. ( A–D ): Induced apoptosis after treatment with varying formulations at 10 h, and the cells were stained using Annexin V-Fluor 647 and 7AAD, then assessed according to manufacturer instructions by the FACScan flow cytometer. UR: Late apoptosis and dead cells; LR: Early apoptosis cells ( A ). PBS; ( B ). Free mitoxantrone; ( C ). mitoxantrone DNA tetrahedra; ( D ). Folate-overhung mitoxantrone DNA tetrahedra; ( E ). The early apoptosis rate in BALL-1 cells ( F ). The late apoptosis rate in BALL-1 cells. ( E–F ): 1. PBS; 2. Free mitoxantrone; 3. Mitoxantrone DNA tetrahedra; 4. Folate-overhung mitoxantrone DNA tetrahedra. a. vs. PBS; b. vs. mitoxantrone DNA tetrahedra, p

    Article Snippet: The fluorescence intensity was measured by the FACScan flow cytometer (Becton Dickinson Biosciences, Franklin Lakes, NJ, USA).

    Techniques: Staining, Flow Cytometry

    Folate deficiency provokes apoptotic lethality in synoviocytes. HIG-82 synoviocytes (1.5×10 5 ) were plated in 60-mm cultured dishes for 24 h. The culture medium was replaced with FC, MFD, and FD media and then continued cultivating for additional 48 h. (A) Cells were then collected, washed with PBS, fixed in PBS-methanol (1:2 v/v) solution and maintained at 4°C for at least 18 h. After one washed with PBS, the cell pellets were then stained with a PI solution containing PBS, PI (40μg/mL), and DNase-free RNase A (40μg/mL) for 30 min at RT in the dark. The cell pellets were then analyzed using a Becton-Dickinson FACSan flowcytometer. The epirubicin (500 nM) treatment (Epi) is a positive control assay of apoptosis. The blank bar, gray bar, right slash bar and left slash bar represent FC, MFD, FD and Epi treatment, respectively. The percentages of subG1 population determined by the PI fluorescent intensity in apoptosis cells which was weaker than that of cells in the G1 phase. The percentages of apoptosis cells were characterized as the percentages of cells in the SubG1 region of the DNA distribution histograms. The FD subG1 bar graph is compared with FC or MFD. A p

    Journal: PLoS ONE

    Article Title: Folate Deficiency Triggered Apoptosis of Synoviocytes: Role of Overproduction of Reactive Oxygen Species Generated via NADPH Oxidase/Mitochondrial Complex II and Calcium Perturbation

    doi: 10.1371/journal.pone.0146440

    Figure Lengend Snippet: Folate deficiency provokes apoptotic lethality in synoviocytes. HIG-82 synoviocytes (1.5×10 5 ) were plated in 60-mm cultured dishes for 24 h. The culture medium was replaced with FC, MFD, and FD media and then continued cultivating for additional 48 h. (A) Cells were then collected, washed with PBS, fixed in PBS-methanol (1:2 v/v) solution and maintained at 4°C for at least 18 h. After one washed with PBS, the cell pellets were then stained with a PI solution containing PBS, PI (40μg/mL), and DNase-free RNase A (40μg/mL) for 30 min at RT in the dark. The cell pellets were then analyzed using a Becton-Dickinson FACSan flowcytometer. The epirubicin (500 nM) treatment (Epi) is a positive control assay of apoptosis. The blank bar, gray bar, right slash bar and left slash bar represent FC, MFD, FD and Epi treatment, respectively. The percentages of subG1 population determined by the PI fluorescent intensity in apoptosis cells which was weaker than that of cells in the G1 phase. The percentages of apoptosis cells were characterized as the percentages of cells in the SubG1 region of the DNA distribution histograms. The FD subG1 bar graph is compared with FC or MFD. A p

    Article Snippet: Measurement of intracellular GSH depletion HIG-82 synoviocytes (1.5×105 ) were plated in 6-cm culture dishes for 24 h. The culture medium was replaced with three types of media: (1) FC medium, (2) MFD medium and (3) FD medium for 48 h. After treatment, cells were incubated with 5 μM CMF-DA for 20 min at 37°C in a 5% CO2 incubator, washed once with PBS, collected by centrifugation, suspended in PBS, and then measured through a 530/22-nm barrier filter using a Becton-Dickinson FACSan flow cytometer.

    Techniques: Cell Culture, Staining, Positive Control Assay

    DPV022 inhibits Bax-induced apoptosis and Bax oligomerization. (A) HeLa cells were cotransfected with pEGFP, pEGFP-DPV022, pEGFP-FPV039, pEGFP-M11L, or pEGFP-Bcl-2 and pcDNA3-HA-Bax. Apoptosis was assessed by quantifying TMRE fluorescence via flow cytometry,

    Journal: Journal of Virology

    Article Title: Deerpox Virus Encodes an Inhibitor of Apoptosis That Regulates Bak and Bax ▿

    doi: 10.1128/JVI.01959-10

    Figure Lengend Snippet: DPV022 inhibits Bax-induced apoptosis and Bax oligomerization. (A) HeLa cells were cotransfected with pEGFP, pEGFP-DPV022, pEGFP-FPV039, pEGFP-M11L, or pEGFP-Bcl-2 and pcDNA3-HA-Bax. Apoptosis was assessed by quantifying TMRE fluorescence via flow cytometry,

    Article Snippet: Cells were analyzed by two-color flow cytometry (FACScan; Becton Dickinson) with TMRE fluorescence measured through the fluorescence parameter 2 (FL-2) channel equipped with a 585-nm filter (42-nm band pass) and EGFP fluorescence measured through the FL-1 channel equipped with a 489-nm filter (42-nm band pass).

    Techniques: Fluorescence, Flow Cytometry, Cytometry

    DPV022 inhibits Bak-induced apoptosis. (A) HeLa cells were cotransfected with pEGFP, pEGFP-DPV022, pEGFP-FPV039, pEGFP-M11L, or pEGFP-Bcl-2 and pcDNA3-HA-Bak. Apoptosis was assessed by quantifying TMRE fluorescence via flow cytometry, and the percentage

    Journal: Journal of Virology

    Article Title: Deerpox Virus Encodes an Inhibitor of Apoptosis That Regulates Bak and Bax ▿

    doi: 10.1128/JVI.01959-10

    Figure Lengend Snippet: DPV022 inhibits Bak-induced apoptosis. (A) HeLa cells were cotransfected with pEGFP, pEGFP-DPV022, pEGFP-FPV039, pEGFP-M11L, or pEGFP-Bcl-2 and pcDNA3-HA-Bak. Apoptosis was assessed by quantifying TMRE fluorescence via flow cytometry, and the percentage

    Article Snippet: Cells were analyzed by two-color flow cytometry (FACScan; Becton Dickinson) with TMRE fluorescence measured through the fluorescence parameter 2 (FL-2) channel equipped with a 585-nm filter (42-nm band pass) and EGFP fluorescence measured through the FL-1 channel equipped with a 489-nm filter (42-nm band pass).

    Techniques: Fluorescence, Flow Cytometry, Cytometry