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Figure 3

Figure 3. From: A high-throughput 3-parameter flow cytometry-based cell death assay.

Flow cytometry-based evaluation of apoptosis. A typical plot of cells treated with A. racemosa extract undergoing apoptosis shows three populations. (A) live cells; (B) early stage apoptosis; and (C) late stage apoptosis. It is also possible to identify the small population of cells that have lost external membrane integrity but still support an intact nuclear membrane (arrow).

Eric J Buenz, et al. Cytometry A. ;71(3):170-173.
FIG. 1.

FIG. 1. From: Comparison of Plaque- and Flow Cytometry-Based Methods for Measuring Dengue Virus Neutralization .

Comparison of flow cytometry and plaque-based DENV neutralization assays. WHO reference dengue immune sera with monospecific immunity to DENV3 or DENV4 were analyzed by the flow cytometry-based neutralization assay with Vero cells (A) and U937 cells expressing DC-SIGN (B) and by PRNT (C). Each serum sample was tested in duplicate or triplicate against the four serotypes in three independent experiments. Dark circles, individual 50% neutralization titer measurements; horizontal lines, the mean titer for each group.

Annette A. Kraus, et al. J Clin Microbiol. 2007 November;45(11):3777-3780.
Figure 2

Figure 2. From: Oxidative stress and PARP activation mediate the NADH-induced decrease in glioma cell survival.

(A) The graphs from flow cytometry-based PI assays showed that NADH treatment induced an increase in the number of PI-positive C6 glioma cells. The P2 fraction and the P3 fraction indicate the number of PI-positive cells and PI-negative cells, respectively. The number of PI-positive cells in the samples treated with 1000 μM NADH is greater than that in the controls. (B) Quantifications of the results from the flow cytometry-based PI assays showed that NADH treatment induced a significant increase in the number of PI-positive C6 glioma cells. The cells were treated with 1000 μM NADH for 24 hrs, and subsequently the number of PI-positive cells was assessed by flow cytometry-based PI assay. N = 9. Data were pooled from 3 independent experiments. ***, p < 0.001.

Yingxin Ma, et al. Int J Physiol Pathophysiol Pharmacol. 2011 January 1;3(1):21-28.
Figure 1

Figure 1. From: Flow Cytometry-Based Methods for Assessing Soluble scFv Activities and Detecting Antigens in Solution.

Schematic diagram of assays employed in this study. The three flow cytometry-based methods for assessing soluble scFv activities and detecting antigens in solution are diagrammed. Positive results are depicted in each case. In CIFC-Ab and CIFC-Ag assays, positive results occur when unlabeled soluble scFv antibodies or unlabeled antigen, respectively, competitively inhibit binding of biotinylated antigen to yeast cells. In the epitope binning assay, a positive result is detected when distinct epitopes are bound by distinct yeast-bound and soluble scFv, resulting in a scFv antibody “sandwich” detectable by appropriate monoclonal antibodies (MAb). SA–PE, streptavidin–phycoerythrin.

Sean A. Gray, et al. Biotechnol Bioeng. ;105(5):973-981.
 Figure 1. 

Figure 1.  . From: Using Flow Cytometry to Compare the Dynamics of Photoreceptor Outer Segment Phagocytosis in iPS-Derived RPE Cells.

(A) Schematic illustrating the flow cytometry–based phagocytosis assay. (B) Electron micrograph showing bound POS at the surface of a murine primary RPE cell (arrow); scale bar = 200 nm. (C) Image showing internalized POS in murine RPE (arrow); scale bar = 1 μm.

Peter D. Westenskow, et al. Invest Ophthalmol Vis Sci. 2012 September;53(10):6282-6290.
Figure 5

Figure 5. High-throughput-screening for unknown protein interactions by FACS-FRET.. From: A Flow Cytometry-Based FRET Assay to Identify and Analyse Protein-Protein Interactions in Living Cells.

(a) Experimental setup to screen for unknown protein interaction with flow cytometry based FRET in high-throughput. (b) Living 293T cells were transfected with the Vpu-YFP fusion as a bait and a mixture of equal amounts of the CFP-fusion constructs that are described in Figure 2. 36 h post transfection FRET+ cells were sorted, pelleted, resuspended in PBS and reanalysed for successful purification. Abbreviation GOI, gene of interest.

Carina Banning, et al. PLoS One. 2010;5(2):e9344.
Fig. 5.2

Fig. 5.2. From: Strategies to Insulate Lentiviral Vector-Expressed Transgenes.

Schematic diagram showing the reporter plasmid pRL-TKGFP-CMVE used in the flow cytometry-based barrier function assay (A) and the reporter plasmid pRL-TKGFP-CMVE-TKRFP used in the flow cytometry-based enhancer blocking assay (B). Abbreviations: 1.2 kb 5′HS4, 1.2-kb fragment of the chicken β-globin 5′HS4 insulator; GFP, enhanced GFP gene; RFP, DsRed.T4 gene; E, human CMV immediate early region enhancer; P, herpes simplex virus TK promoter.

Ali Ramezani, et al. Methods Mol Biol. ;614:77-100.
Figure 7.

Figure 7. From: Notch activation inhibits AML growth and survival: a potential therapeutic approach.

Notch ligand DLL1 co-culture inhibits AML growth, induces apoptosis, down-regulates BCL2, and reveals the critical roles of HES1, BCL2, and p53 in Notch-mediated apoptosis in AML. (A) Cell counts after 72 h of co-culture on human stromal line HS5 transduced with GFP-only or Notch ligand DLL1 GFP. 100,000 AML or T-ALL cells were placed over confluent HS5-GFP or HS5-DLL1 cells. (**, P < 0.001). Triplicate samples analyzed. (B) Flow cytometry–based apoptosis assay in AML and T-ALL cells stained with Annexin V after co-culture with HS5-GFP or HS5-DLL1 cells for 24 h. **, P < 0.001. Triplicate samples were analyzed. (C) Immunoblot of AML and T-ALL cells after co-culture with HS5-GFP or HS5-DLL1 cells for 48 h. Representative of two blots. (D) mRNA expression of BCL2 in AML and T-ALL co-cultured with HS5-GFP or HS5-DLL1 cells for 24 h. **, P < 0.001. Triplicate samples analyzed. (E) Flow cytometry–based apoptosis assay of AML cells transfected with siRNA to HES1 or control, and then co-cultured on HS5-GFP or HS5-DLL1 for 48 h. Representative of two experiments. (F) Immunoblot showing effect of transfection with siRNA to HES1 on level of HES1 protein induced by transduction with ICN1. Representative of two blots. (G) Flow cytometry–based apoptosis assay of AML cells transfected with BCL2, siP53, or BCL2 + siP53 was then co-cultured on HS5-GFP or HS5-DLL1 for 48 h. Representative of two experiments.

Sankaranarayanan Kannan, et al. J Exp Med. 2013 February 11;210(2):321-337.
Figure 2

Figure 2. Induction of neutralizing antibody. From: Adjuvant-Enhanced Antibody Responses to Recombinant Proteins Correlates with Protection of Mice and Monkeys to Orthopoxvirus Challenges.

The sera described in Figure 1 obtained from mice immunized with AL plus the indicated adjuvants were used. MV neutralizing antibodies were measured with a flow cytometry-based GFP assay and the 50% inhibitory concentration (IC50) was determined for each pool of mouse sera

Christiana N. Fogg, et al. Vaccine. ;25(15):2787-2799.
Figure 2

Figure 2. From: Cantharidin and norcantharidin inhibit the ability of MCF-7 cells to adhere to platelets via protein kinase C pathway-dependent downregulation of ?2 integrin.

(A) Photomicrographs and fluorescence microscopy images of adhesion between MCF-7 cells and platelets. (B) Flow cytometry-based platelet adhesion assay. The fluorescent positive rate increased when the platelet concentration increased. (C) Cantharidin (CAN) and norcantharidin (NCTD) treatment decreased the fluorescent positive rate. *P<0.05 vs. control group.

LIU-MEI SHOU, et al. Oncol Rep. 2013 September;30(3):1059-1066.

FIGURE 1:. From: Functional genomic screen identifies novel mediators of collagen uptake.

Flow cytometry–based collagen uptake assay in Drosophila S2. (A–D) S2 cells incubated with FITC-conjugated collagen type I at 4°C (A, C) or 27°C (B, D) for 3 h with the addition of trypan blue (TB) before fluorescence microscopy (A, B) or flow cytometry (C, D).

Ting-Hein Lee, et al. Mol Biol Cell. 2014 March 1;25(5):583-593.
Figure 4

Figure 4. From: IL-15, in synergy with RAE-1?, stimulates TCR-independent proliferation and activation of CD8+ T cells.

Lysis activity of BaF3-mb15-RAE-stimulated CD8+ T cells. CFSE-labeled target cells were co-incubated with CD8+ T cells at E:T ratios of 2.5:1 to 20:1 in the presence of the indicated irradiated stimulatory cells. The specific lysis was determined by a flow cytometry-based cytotoxicity assay. Results are presented as the means ± SD of triplicates. E:T, effector-target ratio.

LI QIAN, et al. Oncol Lett. 2012 February;3(2):472-476.
Figure 2

Figure 2. From: Anti-lung cancer effects of novel ginsenoside 25-OCH3-PPD.

(A) Anti-proliferative effects of 25-OCH3-PPD on lung cancer cells, A549, H358, and H838, in comparison with control cell line BEAS-2B. These cells were exposed to various concentrations of the ginsenosides for 24 h followed by the BrdU incorporation assay. (B) Induction of apoptosis by 25-OCH3-PPD in A549, H358, and H838 lung cancer cells, in comparison with the control cell line BEAS-2B. Cells were exposed to various concentrations of the ginsenosides for 48 h followed by the flow cytometry-based apoptosis assay. All assays were performed in triplicate.

Wei Wang, et al. Lung Cancer. ;65(3):306-311.
Figure 4.

Figure 4. From: Notch activation inhibits AML growth and survival: a potential therapeutic approach.

The Notch target gene HES1 induces growth arrest via its repressor domain. (A) Flow cytometry–based competitive proliferation assay showing changes in GFP after HES1 transduction in seven AML lines (mean ± SD). Triplicate samples were analyzed. (B) Flow cytometry–based assay showing changes in GFP in cells stably transduced with a tamoxifen-inducible ER-HES1 and exposed to 4-OH tamoxifen at the doses and the time durations indicated. Representative of two experiments. (C) Flow cytometry–based cell cycle histogram showing DNA content measured by propidium iodide (PI) in ER-HES1 cells exposed to 4 µM 4-OH tamoxifen for 48 h. Representative of three experiments. (D) HES1 mutants lacking either the conserved WRPW motif (HES1-GRPG) or the C-terminal repressor domain (HES1-deltaC) were constructed. (E) HES1 mutants were transduced into AML cells. A competitive proliferation assay reveals growth effects by wild-type HES1, the GRPG mutant, and the HES1-deltaC mutant (mean ± SD). **, P < 0.001. Triplicate samples analyzed and similar results were obtained in K562 and HL60 lines.

Sankaranarayanan Kannan, et al. J Exp Med. 2013 February 11;210(2):321-337.
Figure 1

Figure 1. From: Aldehyde dehydrogenase activity is a cancer stem cell marker of tongue squamous cell carcinoma.

ALDH+ cells detected in the Tca8113 tongue squamous cell carcinoma cell line. ALDH+ Tca8113 cells were identified using a flow cytometry-based Aldefluor assay. (A) Baseline fluorescence was established in the presence of ALDH inhibitor DEAB. (B) ALDH+ cells detected in Tca8113 cells without DEAB.

BO ZOU, et al. Mol Med Rep. 2012 April;5(4):1116-1120.
Figure 4

Figure 4. From: A flow cytometry based assay to assess RSV specific neutralizing antibody is reproducible, efficient and accurate.

Comparison of plaque reduction neutralization test and flow cytometry neutralization assay. 68 cotton rat serum samples were tested by plaque reduction neutralization test and flow cytometry neutralization. The linear regression analysis demonstrate significant correlation, R2= 0.8444, P< 0.0001.

M Chen, et al. J Immunol Methods. ;362(1-2):180-184.

FIGURE 2. From: Mutations Define Cross-talk between the N-terminal Nucleotide-binding Domain and Transmembrane Helix-2 of the Yeast Multidrug Transporter Pdr5.

R6G transport is drastically diminished in S558Y. The transport activity of WT and mutant Pdr5s was estimated with a flow cytometry-based efflux assay with R6G as the fluorescent substrate. The null strain (Δpdr5 = AD1–7) and yeast strains containing the WT (JG2000) and mutant (JG2009) Pdr5 were used in these experiments. The assay was carried out as described previously (21), and the representative histograms from four independent experiments are depicted. The strains used are labeled on the figure.

Zuben E. Sauna, et al. J Biol Chem. 2008 December 12;283(50):35010-35022.
Fig 9

Fig 9. From: Persistence of Epstein-Barr Virus in Self-Reactive Memory B Cells.

Reactivity of antibodies derived from donor-matched EBV+ and EBV memory B cells to MOG. Reactivity was assessed with a flow cytometry-based assay using an oligodendrocyte cell line expressing MOG. Positive (anti-MOG antibody)- and negative (irrelevant antibody)-control (ctrl) antibody binding is shown by the gray and black bars, respectively. Reactivity is expressed as the ratio of the MFI of staining on MOG+ cells to the MFI of staining on MOG cells.

Sean I. Tracy, et al. J Virol. 2012 November;86(22):12330-12340.
Figure 3

Figure 3. Cytokine concentrations of Basal Tears (BT), Reflex tears obtained in Sitting position (RS) and Reflex tears collected with the subject Lying (RL) in Normal Control (NC) eyes.. From: Human Tears Reveal Insights into Corneal Neovascularization.

Tears of 10 control subjects were analysed for cytokine secretion using a flow cytometry based assay. Bars indicate mean concentrations and differences are significant if *p<0.05 **p<0.001, ***p<0.0001.

Nadia Zakaria, et al. PLoS One. 2012;7(5):e36451.
Figure 4

Figure 4. From: In vitro micronucleus assay scored by flow cytometry provides a comprehensive evaluation of cytogenetic damage and cytotoxicity.

Micronucleus data are graphed for each of six test agents studied. Mean micronucleus responses (fold-increase values) are shown for microscopy-based measurements (white bars) and also for flow cytometry-based assessments (black bars). Asterisks indicate average fold-increases ≥ 3x over negative control. All dose levels that exhibited ≥ 40% relative survival as measured by Coulter counter are shown here. Doses that were deemed inappropriately high based on flow cytometric analyses are indicated by a bracket and the word “cytotoxic”.

Steven M. Bryce, et al. Mutat Res. ;630(1-2):78-91.

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