Structured Review

BioTek Instruments microplate reader
High lytic activity of ClyR. ( A ) Time-kill curves of ClyR against S. dysgalactiae ATCC 35666. Bacterial cells were washed once with PBS, treated with 25 μg/ml ClyR, and the change of OD 600 (right Y-axis) were monitored by a <t>microplate</t> reader at 37 °C for 30 min. Triangles: ClyR; Circles: PBS controls. In parallel, the viable cell numbers (squares, left Y-axis) were calculated by plating onto BHI agar plates at different time points. ( B ) Dose-dependent lytic efficacy of ClyR against S. agalactiae S12, S. dysgalactiae ATCC 35666, or an equal mixture of both strains in market pasteurized milk at 30 °C for 1 h. ( C ) TEM images of S. dysgalactiae ATCC 35666 cells exposed to ClyR. Bar sizes: 500 nm. ( D ) Time-killing efficacy of ClyR (40 μg/ml) against S. agalactiae S12, S. dysgalactiae ATCC 35666, or an equal mixture of both strains in market pasteurized milk at 30 °C. ( E ) Lytic efficacy of ClyR (40 μg/ml) in pasteurized cow’s milk against S. agalactiae S12, or S. dysgalactiae ATCC 35666 at 30 °C for 1 h. Fresh cow milk samples A, B, and C were taken from healthy cows, and samples D and E were from mastitic cows. ( F ) Comparison of the activity of 0.89 μM ClyR or PlyCAC against various strains. ( G ) Comparison of the activity of 0.89 μM ClyR or PlyGBS-180 against S. dysgalactiae ATCC 35666. ( H ) Efficacy of PlyGBS-180 against S. dysgalactiae ATCC 35666 in market pasteurized milk at 30 °C for 60 min.
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Images

1) Product Images from "A chimeolysin with extended-spectrum streptococcal host range found by an induced lysis-based rapid screening method"

Article Title: A chimeolysin with extended-spectrum streptococcal host range found by an induced lysis-based rapid screening method

Journal: Scientific Reports

doi: 10.1038/srep17257

High lytic activity of ClyR. ( A ) Time-kill curves of ClyR against S. dysgalactiae ATCC 35666. Bacterial cells were washed once with PBS, treated with 25 μg/ml ClyR, and the change of OD 600 (right Y-axis) were monitored by a microplate reader at 37 °C for 30 min. Triangles: ClyR; Circles: PBS controls. In parallel, the viable cell numbers (squares, left Y-axis) were calculated by plating onto BHI agar plates at different time points. ( B ) Dose-dependent lytic efficacy of ClyR against S. agalactiae S12, S. dysgalactiae ATCC 35666, or an equal mixture of both strains in market pasteurized milk at 30 °C for 1 h. ( C ) TEM images of S. dysgalactiae ATCC 35666 cells exposed to ClyR. Bar sizes: 500 nm. ( D ) Time-killing efficacy of ClyR (40 μg/ml) against S. agalactiae S12, S. dysgalactiae ATCC 35666, or an equal mixture of both strains in market pasteurized milk at 30 °C. ( E ) Lytic efficacy of ClyR (40 μg/ml) in pasteurized cow’s milk against S. agalactiae S12, or S. dysgalactiae ATCC 35666 at 30 °C for 1 h. Fresh cow milk samples A, B, and C were taken from healthy cows, and samples D and E were from mastitic cows. ( F ) Comparison of the activity of 0.89 μM ClyR or PlyCAC against various strains. ( G ) Comparison of the activity of 0.89 μM ClyR or PlyGBS-180 against S. dysgalactiae ATCC 35666. ( H ) Efficacy of PlyGBS-180 against S. dysgalactiae ATCC 35666 in market pasteurized milk at 30 °C for 60 min.
Figure Legend Snippet: High lytic activity of ClyR. ( A ) Time-kill curves of ClyR against S. dysgalactiae ATCC 35666. Bacterial cells were washed once with PBS, treated with 25 μg/ml ClyR, and the change of OD 600 (right Y-axis) were monitored by a microplate reader at 37 °C for 30 min. Triangles: ClyR; Circles: PBS controls. In parallel, the viable cell numbers (squares, left Y-axis) were calculated by plating onto BHI agar plates at different time points. ( B ) Dose-dependent lytic efficacy of ClyR against S. agalactiae S12, S. dysgalactiae ATCC 35666, or an equal mixture of both strains in market pasteurized milk at 30 °C for 1 h. ( C ) TEM images of S. dysgalactiae ATCC 35666 cells exposed to ClyR. Bar sizes: 500 nm. ( D ) Time-killing efficacy of ClyR (40 μg/ml) against S. agalactiae S12, S. dysgalactiae ATCC 35666, or an equal mixture of both strains in market pasteurized milk at 30 °C. ( E ) Lytic efficacy of ClyR (40 μg/ml) in pasteurized cow’s milk against S. agalactiae S12, or S. dysgalactiae ATCC 35666 at 30 °C for 1 h. Fresh cow milk samples A, B, and C were taken from healthy cows, and samples D and E were from mastitic cows. ( F ) Comparison of the activity of 0.89 μM ClyR or PlyCAC against various strains. ( G ) Comparison of the activity of 0.89 μM ClyR or PlyGBS-180 against S. dysgalactiae ATCC 35666. ( H ) Efficacy of PlyGBS-180 against S. dysgalactiae ATCC 35666 in market pasteurized milk at 30 °C for 60 min.

Techniques Used: Activity Assay, Transmission Electron Microscopy

2) Product Images from "Paracoccin Induces M1 Polarization of Macrophages via Interaction with TLR4"

Article Title: Paracoccin Induces M1 Polarization of Macrophages via Interaction with TLR4

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2016.01003

Purification and biological activities of a recombinant form of paracoccin produced using the Pichia pastoris expression system. (A) SDS-PAGE of the fraction of the culture supernatant that was adsorbed to chitin and eluted in acidic buffer; shows a 27 kDa silver stained band, as determined on the basis of the migration index of MM markers (lane MW). (B) Binding of the purified recombinant paracoccin (named p-rPCN, 200 ng) to the laminin coated wells (250 ng) of a microplate. The binding was revealed by reaction with anti-paracoccin IgY. (C) N -acetyl-glucosaminidase activity exerted by the p-rPCN as revealed using the substrate ρ-nitrophenyl- N -acetyl-β- D -glucosaminide (100 μL 5 mM ρ-NPGlcNAc). The results are expressed as mean ± SEM and were compared to the medium through one-way analysis of variance, followed by Bonferroni’s test. ∗∗∗ p
Figure Legend Snippet: Purification and biological activities of a recombinant form of paracoccin produced using the Pichia pastoris expression system. (A) SDS-PAGE of the fraction of the culture supernatant that was adsorbed to chitin and eluted in acidic buffer; shows a 27 kDa silver stained band, as determined on the basis of the migration index of MM markers (lane MW). (B) Binding of the purified recombinant paracoccin (named p-rPCN, 200 ng) to the laminin coated wells (250 ng) of a microplate. The binding was revealed by reaction with anti-paracoccin IgY. (C) N -acetyl-glucosaminidase activity exerted by the p-rPCN as revealed using the substrate ρ-nitrophenyl- N -acetyl-β- D -glucosaminide (100 μL 5 mM ρ-NPGlcNAc). The results are expressed as mean ± SEM and were compared to the medium through one-way analysis of variance, followed by Bonferroni’s test. ∗∗∗ p

Techniques Used: Purification, Recombinant, Produced, Expressing, SDS Page, Staining, Migration, Binding Assay, Activity Assay

3) Product Images from "An in vitro study on sonodynamic treatment of human colon cancer cells using sinoporphyrin sodium as sonosensitizer"

Article Title: An in vitro study on sonodynamic treatment of human colon cancer cells using sinoporphyrin sodium as sonosensitizer

Journal: BioMedical Engineering OnLine

doi: 10.1186/s12938-020-00797-w

Intracellular uptake of DVDMS. a Intracellular localization of DVDMS (red) in HCT116 cells and RKO cells. The nuclei were stained with DAPI (blue). Scale bar 50 µm. b Measurement of fluorescence intensity of intracellular DVDMS in HCT116, RKO and NCM460 cells with different incubation durations by a microplate reader and flow cytometry
Figure Legend Snippet: Intracellular uptake of DVDMS. a Intracellular localization of DVDMS (red) in HCT116 cells and RKO cells. The nuclei were stained with DAPI (blue). Scale bar 50 µm. b Measurement of fluorescence intensity of intracellular DVDMS in HCT116, RKO and NCM460 cells with different incubation durations by a microplate reader and flow cytometry

Techniques Used: Staining, Fluorescence, Incubation, Flow Cytometry

4) Product Images from "Metabolic Flux Analysis of Catechin Biosynthesis Pathways Using Nanosensor"

Article Title: Metabolic Flux Analysis of Catechin Biosynthesis Pathways Using Nanosensor

Journal: Antioxidants

doi: 10.3390/antiox9040288

Specificity analysis of FLIP-Cat. Purified nanosensor protein (20 mg/mL) was taken in different wells of a microplate and various homologous metabolites of catechin (0, 1, and 10 mM) were added. In one well (+)-catechin (0, 1, and 10 mM) was added. The fluorescence resonance energy transfer (FRET) ratio was recorded. Values are the means of three independent replicate. Vertical bars show the standard error.
Figure Legend Snippet: Specificity analysis of FLIP-Cat. Purified nanosensor protein (20 mg/mL) was taken in different wells of a microplate and various homologous metabolites of catechin (0, 1, and 10 mM) were added. In one well (+)-catechin (0, 1, and 10 mM) was added. The fluorescence resonance energy transfer (FRET) ratio was recorded. Values are the means of three independent replicate. Vertical bars show the standard error.

Techniques Used: Purification, Fluorescence, Förster Resonance Energy Transfer

5) Product Images from "Triangular correlation (TrC) between cancer aggressiveness, cell uptake capability, and cell deformability"

Article Title: Triangular correlation (TrC) between cancer aggressiveness, cell uptake capability, and cell deformability

Journal: Science Advances

doi: 10.1126/sciadv.aax2861

Cancer skin cells (A375-P) present an enhanced uptake compared with noncancer skin cells (HaCaT). ( A ) FACS analysis is presented as column plot of normalized average uptake values of 0.05-, 0.3-, 0.5-, 0.8-, and 2.4-μm fluorescently labeled polystyrene beads. Representative histograms for 2.4-μm beads are shown at the bottom. ( B ) Microplate reader fluorescence analysis. Excitation, 530/25 and emission, 590/20, with 0.3-, 0.8-, 2.4-, 6-, 10.4-, and 15.7-μm beads. The unitless results in the column plots of (A) and (B) are normalized by the intensity of particle fluorescence to represent the relative mass of cell internalized particles and are scaled to a maximal value of 1 (see eq. S1). ( C ) Fluorescent microscopy images of cells incubated with 0.3-, 0.8-, and 2.4-μm beads. Blue, DAPI; red, beads. Scale bars, 10 μm (upper panels, 0.3 μm) or 50 μm (middle and lower panels, 0.8 and 2.4 μm).
Figure Legend Snippet: Cancer skin cells (A375-P) present an enhanced uptake compared with noncancer skin cells (HaCaT). ( A ) FACS analysis is presented as column plot of normalized average uptake values of 0.05-, 0.3-, 0.5-, 0.8-, and 2.4-μm fluorescently labeled polystyrene beads. Representative histograms for 2.4-μm beads are shown at the bottom. ( B ) Microplate reader fluorescence analysis. Excitation, 530/25 and emission, 590/20, with 0.3-, 0.8-, 2.4-, 6-, 10.4-, and 15.7-μm beads. The unitless results in the column plots of (A) and (B) are normalized by the intensity of particle fluorescence to represent the relative mass of cell internalized particles and are scaled to a maximal value of 1 (see eq. S1). ( C ) Fluorescent microscopy images of cells incubated with 0.3-, 0.8-, and 2.4-μm beads. Blue, DAPI; red, beads. Scale bars, 10 μm (upper panels, 0.3 μm) or 50 μm (middle and lower panels, 0.8 and 2.4 μm).

Techniques Used: FACS, Labeling, Fluorescence, Microscopy, Incubation

Uptake of beads by cancer cells has a nonmonotonic dependence on the bead size. Cancer cell interaction with inert spherical fluorescent polystyrene beads increases with bead size, reaching a maximum around 2.4-μm bead diameter and then decreases. ( A ) Left: FACS analysis; average over normalized uptake values of eight cell lines: A375-p, A375-SM, PC3M-P, PC3M, LN4253J, 253 J-B5, SN12, and SN12-C. Right: Microplate fluorescent analysis (excitation, 530/25 and emission, 590/20), average over normalized uptake values of A375-P, A375-SM, PC3M-P, and PC3M-LN4. ( B ) Confocal fluorescent images (blue, DAPI; green, fluorescein isothiocyanate–phalloidin; red, polystyrene fluorescently labeled beads). Scale bars, 10 μm. ( C ) Physical model predicted the nonmonotonic dependence due to the interplay between adhesion and cell deformability. ( D ) The free energy of the wrapping model has a minimum as a function of both the bead radius and the wrapping angle, ϑ (bottom); however, for small beads of R
Figure Legend Snippet: Uptake of beads by cancer cells has a nonmonotonic dependence on the bead size. Cancer cell interaction with inert spherical fluorescent polystyrene beads increases with bead size, reaching a maximum around 2.4-μm bead diameter and then decreases. ( A ) Left: FACS analysis; average over normalized uptake values of eight cell lines: A375-p, A375-SM, PC3M-P, PC3M, LN4253J, 253 J-B5, SN12, and SN12-C. Right: Microplate fluorescent analysis (excitation, 530/25 and emission, 590/20), average over normalized uptake values of A375-P, A375-SM, PC3M-P, and PC3M-LN4. ( B ) Confocal fluorescent images (blue, DAPI; green, fluorescein isothiocyanate–phalloidin; red, polystyrene fluorescently labeled beads). Scale bars, 10 μm. ( C ) Physical model predicted the nonmonotonic dependence due to the interplay between adhesion and cell deformability. ( D ) The free energy of the wrapping model has a minimum as a function of both the bead radius and the wrapping angle, ϑ (bottom); however, for small beads of R

Techniques Used: FACS, Labeling

6) Product Images from "Light-assisted small molecule screening against protein kinases"

Article Title: Light-assisted small molecule screening against protein kinases

Journal: Nature chemical biology

doi: 10.1038/nchembio.1933

Optogenetics-enabled, internally-referenced measurement of MAPK/ERK pathway ( a ) Whole well (I.) or spatially-confined (II.) light stimulation (λ=470 ± 5 nm) of SPC212 Opto-mFGFR1 cells was performed in a microplate reader. 48-well plates were chosen for these experiments to enable visual evaluation after anti -pERK1/2 immunohistochemistry (see b and d). ( b ) Raw data photographs of cells stimulated with EGF (5.5 ng/ml) or light (distributed over the well in a 3×3 matrix). MAPK/ERK pathway was activated by EGF in SPC212 cells or light in SPC212 Opto-mFGFR1 cells but not in controls. ( c ) Quantification of b. Mean (normalized) absorption values ± SEM (n=9, one representative experiment) are shown. ( d ) Raw data photographs of local activation (area~3.14 mm 2 ) of the MAPK/ERK pathway by spatially-confined illumination of SPC212 Opto-mFGFR1 cells. Activation is limited to the center of the well and inhibited by PD-166866 (final concentration 5 μM). ( e ) Quantification of d. ( f ) Characterization of an inactive molecule (vehicle, left bars), inhibitor (PD-166866, middle bars) or activator (EGF, right bars) through internal references in a single measurement. Mean (normalized) absorption values ± SEM (n=6, 3 and 2 from left to right, two independent experiments) are shown. Scale bar in b and c is 1 mm.
Figure Legend Snippet: Optogenetics-enabled, internally-referenced measurement of MAPK/ERK pathway ( a ) Whole well (I.) or spatially-confined (II.) light stimulation (λ=470 ± 5 nm) of SPC212 Opto-mFGFR1 cells was performed in a microplate reader. 48-well plates were chosen for these experiments to enable visual evaluation after anti -pERK1/2 immunohistochemistry (see b and d). ( b ) Raw data photographs of cells stimulated with EGF (5.5 ng/ml) or light (distributed over the well in a 3×3 matrix). MAPK/ERK pathway was activated by EGF in SPC212 cells or light in SPC212 Opto-mFGFR1 cells but not in controls. ( c ) Quantification of b. Mean (normalized) absorption values ± SEM (n=9, one representative experiment) are shown. ( d ) Raw data photographs of local activation (area~3.14 mm 2 ) of the MAPK/ERK pathway by spatially-confined illumination of SPC212 Opto-mFGFR1 cells. Activation is limited to the center of the well and inhibited by PD-166866 (final concentration 5 μM). ( e ) Quantification of d. ( f ) Characterization of an inactive molecule (vehicle, left bars), inhibitor (PD-166866, middle bars) or activator (EGF, right bars) through internal references in a single measurement. Mean (normalized) absorption values ± SEM (n=6, 3 and 2 from left to right, two independent experiments) are shown. Scale bar in b and c is 1 mm.

Techniques Used: Optogenetics, Immunohistochemistry, Activation Assay, Concentration Assay

7) Product Images from "GSH-sensitive Pt(IV) prodrug-loaded phase-transitional nanoparticles with a hybrid lipid-polymer shell for precise theranostics against ovarian cancer"

Article Title: GSH-sensitive Pt(IV) prodrug-loaded phase-transitional nanoparticles with a hybrid lipid-polymer shell for precise theranostics against ovarian cancer

Journal: Theranostics

doi: 10.7150/thno.29820

Mitochondrial apoptosis signaling pathways. (A) Mitochondrial membrane potential (Δ ψ m ) in cells with the treatment of NP-cRGD+US, cisplatin, Pt(IV), Pt(IV) NPs, Pt(IV) NPs+US, Pt(IV) NP-cRGD and Pt(IV) NP-cRGD+US (30 μM eq.) for 48 h, as detected by the ratio of fluorescent intensity ([green]/[red]) of JC-1. (B) Immunohistochemically stained images of cytochrome c translocated from the mitochondria to the cytosol. Original magnification, 200×; Scale bar, 100 μm. (C, D) Activities of caspase 9 and caspase 3 as determined by a microplate reader. (E) Western blotting analyses of Bcl-2, Bax, Cleaved caspase 9 and Cytochrome c after different drug formulations incubations. (F, G) Cell apoptosis of the SKOV3 cells treated with NP-cRGD+US, cisplatin, Pt(IV), Pt(IV) NPs, Pt(IV) NPs+US, Pt(IV) NP-cRGD and Pt(IV) NP-cRGD+US (30 μM eq.) for 48 h, using the Annexin V and PI kit to detect the apoptosis ratio through the flow cytometry analysis, along with the statistics of cell apoptosis ratio, EA: early apoptosis; LA: late apoptosis; TA: total apoptosis. The data are presented as the mean ± SD of three independent experiments. Statistical significance in (A) , (C) and (D) was calculated by one-way ANOVA with Tukey's post hoc test. Statistical significance in (G) was calculated by two-way ANOVA with Tukey's post hoc test. * P
Figure Legend Snippet: Mitochondrial apoptosis signaling pathways. (A) Mitochondrial membrane potential (Δ ψ m ) in cells with the treatment of NP-cRGD+US, cisplatin, Pt(IV), Pt(IV) NPs, Pt(IV) NPs+US, Pt(IV) NP-cRGD and Pt(IV) NP-cRGD+US (30 μM eq.) for 48 h, as detected by the ratio of fluorescent intensity ([green]/[red]) of JC-1. (B) Immunohistochemically stained images of cytochrome c translocated from the mitochondria to the cytosol. Original magnification, 200×; Scale bar, 100 μm. (C, D) Activities of caspase 9 and caspase 3 as determined by a microplate reader. (E) Western blotting analyses of Bcl-2, Bax, Cleaved caspase 9 and Cytochrome c after different drug formulations incubations. (F, G) Cell apoptosis of the SKOV3 cells treated with NP-cRGD+US, cisplatin, Pt(IV), Pt(IV) NPs, Pt(IV) NPs+US, Pt(IV) NP-cRGD and Pt(IV) NP-cRGD+US (30 μM eq.) for 48 h, using the Annexin V and PI kit to detect the apoptosis ratio through the flow cytometry analysis, along with the statistics of cell apoptosis ratio, EA: early apoptosis; LA: late apoptosis; TA: total apoptosis. The data are presented as the mean ± SD of three independent experiments. Statistical significance in (A) , (C) and (D) was calculated by one-way ANOVA with Tukey's post hoc test. Statistical significance in (G) was calculated by two-way ANOVA with Tukey's post hoc test. * P

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

8) Product Images from "Inhibition of Inflammatory Response by Artepillin C in Activated RAW264.7 Macrophages"

Article Title: Inhibition of Inflammatory Response by Artepillin C in Activated RAW264.7 Macrophages

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2013/735176

Effect of artepillin C on chemiluminescence of PMA activated RAW264.7 macrophages: (a) time course of chemiluminescence, (b) chemiluminescence of PMA activated RAW264.7 macrophages treated with 1–50 μ M artepillin C for 30 min. Chemiluminescence was determined using microplate luminometer and expressed as a percentage of PMA-stimulated cells. The values represent mean ± SD of four independent experiments ( n = 8) *** P
Figure Legend Snippet: Effect of artepillin C on chemiluminescence of PMA activated RAW264.7 macrophages: (a) time course of chemiluminescence, (b) chemiluminescence of PMA activated RAW264.7 macrophages treated with 1–50 μ M artepillin C for 30 min. Chemiluminescence was determined using microplate luminometer and expressed as a percentage of PMA-stimulated cells. The values represent mean ± SD of four independent experiments ( n = 8) *** P

Techniques Used:

9) Product Images from "The Static Magnetic Field Remotely Boosts the Efficiency of Doxorubicin through Modulating ROS Behaviors"

Article Title: The Static Magnetic Field Remotely Boosts the Efficiency of Doxorubicin through Modulating ROS Behaviors

Journal: Scientific Reports

doi: 10.1038/s41598-018-19247-8

The total intracellular glutathione (tGSH) results of ( a ) MCF-7, ( b ) HFF cells and ( c ) both comparison exposed with 10 mT SMF and incubated with 0.1 μM DOXO for two exposure times (24 and 48 h). The GSH was measured by a microplate reader, and results are expressed as the concentration of glutathione content (μM GSH per 10 6 cells). Data are shown as mean ± SD (n = 3). **P
Figure Legend Snippet: The total intracellular glutathione (tGSH) results of ( a ) MCF-7, ( b ) HFF cells and ( c ) both comparison exposed with 10 mT SMF and incubated with 0.1 μM DOXO for two exposure times (24 and 48 h). The GSH was measured by a microplate reader, and results are expressed as the concentration of glutathione content (μM GSH per 10 6 cells). Data are shown as mean ± SD (n = 3). **P

Techniques Used: Incubation, Concentration Assay

10) Product Images from "Whitening Effect of Watersoluble Royal Jelly from South Korea"

Article Title: Whitening Effect of Watersoluble Royal Jelly from South Korea

Journal: Korean Journal for Food Science of Animal Resources

doi: 10.5851/kosfa.2015.35.5.707

Cell viability after WSRJ in B16F1 cells. B16F1 cells were treated with 10 nM α-MSH for 48 h and then further 24 h with WSRJ at 1-100 mg/mL. Cell viability was determined by measuring the absorbance at 570 nm using a microplate reader. Data are presented as mean±SEM of five independent experiments. Different letters indicate a significant difference with p
Figure Legend Snippet: Cell viability after WSRJ in B16F1 cells. B16F1 cells were treated with 10 nM α-MSH for 48 h and then further 24 h with WSRJ at 1-100 mg/mL. Cell viability was determined by measuring the absorbance at 570 nm using a microplate reader. Data are presented as mean±SEM of five independent experiments. Different letters indicate a significant difference with p

Techniques Used:

11) Product Images from "Heme-binding-mediated negative regulation of the tryptophan metabolic enzyme indoleamine 2,3-dioxygenase 1 (IDO1) by IDO2"

Article Title: Heme-binding-mediated negative regulation of the tryptophan metabolic enzyme indoleamine 2,3-dioxygenase 1 (IDO1) by IDO2

Journal: Experimental & Molecular Medicine

doi: 10.1038/emm.2014.69

Alleviation of cell growth arrest and death by co-expression of hIDO2. ( a ) Morphological changes in hIDO1-, hIDO2- or hIDO1-2-expressing HEK293 stable cells in the absence or presence of 25 μ M L -Trp were detected on day 3 during the cell proliferation assay ( × 100 magnification). Cell proliferation was measured by WST colorimetric assay during the 3-day culture period in the ( b ) absence or ( c ) presence of 25 μ M L -Trp. Absorbance at 450 nm was measured using a microplate reader. Cell proliferation values on day 2 were subjected to statistical analysis. All experiments were performed independently three times. ns, not significant, *** P
Figure Legend Snippet: Alleviation of cell growth arrest and death by co-expression of hIDO2. ( a ) Morphological changes in hIDO1-, hIDO2- or hIDO1-2-expressing HEK293 stable cells in the absence or presence of 25 μ M L -Trp were detected on day 3 during the cell proliferation assay ( × 100 magnification). Cell proliferation was measured by WST colorimetric assay during the 3-day culture period in the ( b ) absence or ( c ) presence of 25 μ M L -Trp. Absorbance at 450 nm was measured using a microplate reader. Cell proliferation values on day 2 were subjected to statistical analysis. All experiments were performed independently three times. ns, not significant, *** P

Techniques Used: Expressing, Proliferation Assay, Colorimetric Assay

Effects of the indoleamine 2,3-dioxygenase 1 (IDO1) inhibitor L -1MT on human (h) IDO1 activity in cells co-expressing hIDO2 (H) mutants. ( a ) Kynurenine production was measured in culture supernatants of stable hIDO1-expressing cells transfected with 2 μg hIDO2 or hIDO2 (H) mutant DNAs. Transfected cells were incubated for 24 h with complete medium including 100 μ M L -Trp in the absence or presence of 100 μ M L -1MT. ( b ) Proliferation of stable hIDO1-, hIDO1-2- and hIDO1-2 (H)-expressing HEK293 cells was measured by WST assay during the 3-day culture period. Cell proliferation values on day 2 were subjected to statistical analysis. Absorbance at 450 nm was measured using a microplate reader with the Gen5 software. ns, not significant, * P
Figure Legend Snippet: Effects of the indoleamine 2,3-dioxygenase 1 (IDO1) inhibitor L -1MT on human (h) IDO1 activity in cells co-expressing hIDO2 (H) mutants. ( a ) Kynurenine production was measured in culture supernatants of stable hIDO1-expressing cells transfected with 2 μg hIDO2 or hIDO2 (H) mutant DNAs. Transfected cells were incubated for 24 h with complete medium including 100 μ M L -Trp in the absence or presence of 100 μ M L -1MT. ( b ) Proliferation of stable hIDO1-, hIDO1-2- and hIDO1-2 (H)-expressing HEK293 cells was measured by WST assay during the 3-day culture period. Cell proliferation values on day 2 were subjected to statistical analysis. Absorbance at 450 nm was measured using a microplate reader with the Gen5 software. ns, not significant, * P

Techniques Used: Activity Assay, Expressing, Transfection, Mutagenesis, Incubation, WST Assay, Software

12) Product Images from "Myricitrin Attenuates High Glucose-Induced Apoptosis through Activating Akt-Nrf2 Signaling in H9c2 Cardiomyocytes"

Article Title: Myricitrin Attenuates High Glucose-Induced Apoptosis through Activating Akt-Nrf2 Signaling in H9c2 Cardiomyocytes

Journal: Molecules

doi: 10.3390/molecules21070880

The effects of myricitrin on HG-induced ROS production and MMP (ΔΨ m ) reduction in H9c2 cells. ( A ) Fluorescence images and bar diagram showed the ROS levels in the H9c2 cells; the fluorescence intensity of ROS was measured by a fluorescence microplate reader. The bar represents 200 μm; ( B ) Representative images and quantitative analysis of JC-1 staining. Treating H9c2 cells with HG caused a significant decrease in the in the ratio of red to green fluorescence intensity, which is a sign of the early stages of cell apoptosis. The bar represents 400 μm. Values are represented as the mean ± SD; n = 10 wells per group. * p
Figure Legend Snippet: The effects of myricitrin on HG-induced ROS production and MMP (ΔΨ m ) reduction in H9c2 cells. ( A ) Fluorescence images and bar diagram showed the ROS levels in the H9c2 cells; the fluorescence intensity of ROS was measured by a fluorescence microplate reader. The bar represents 200 μm; ( B ) Representative images and quantitative analysis of JC-1 staining. Treating H9c2 cells with HG caused a significant decrease in the in the ratio of red to green fluorescence intensity, which is a sign of the early stages of cell apoptosis. The bar represents 400 μm. Values are represented as the mean ± SD; n = 10 wells per group. * p

Techniques Used: Fluorescence, Staining

13) Product Images from "A colorimetric assay for vanillin detection by determination of the luminescence of o-toluidine condensates"

Article Title: A colorimetric assay for vanillin detection by determination of the luminescence of o-toluidine condensates

Journal: PLoS ONE

doi: 10.1371/journal.pone.0194010

Determination of vanillin standard samples by microplate reader
Figure Legend Snippet: Determination of vanillin standard samples by microplate reader

Techniques Used:

Determination of vanillin standard samples by microplate reader
Figure Legend Snippet: Determination of vanillin standard samples by microplate reader

Techniques Used:

14) Product Images from "Myosin-1a powers the sliding of apical membrane along microvillar actin bundles"

Article Title: Myosin-1a powers the sliding of apical membrane along microvillar actin bundles

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200701144

Quantitative assay for BB membrane shedding. (A) Schematic of the assay used to quantify the extent of BB membrane shedding under various conditions. BBs are labeled with AM1-43, reacted with 2 mM ATP, centrifuged to isolate the shed membrane (5,000- g supernatant), and the amount fluorescent material is measured in a microplate reader. (B) Laser scanning confocal micrograph of phalloidin-labeled (green) isolated BBs shows that AM1-43 (red) evenly labels the BB membrane. Bars, 4 μm. (C) Increasing concentrations of BBs were treated with 2 mM ATP and the amount of membrane shed from each sample was measured using the microplate assay. ATP-induced membrane shedding scales with the BB concentration, demonstrating a linear response over two orders of magnitude. Points on this plot represent mean ± SD calculated from replicates at each BB concentration (for all values, SD is smaller than the point size).
Figure Legend Snippet: Quantitative assay for BB membrane shedding. (A) Schematic of the assay used to quantify the extent of BB membrane shedding under various conditions. BBs are labeled with AM1-43, reacted with 2 mM ATP, centrifuged to isolate the shed membrane (5,000- g supernatant), and the amount fluorescent material is measured in a microplate reader. (B) Laser scanning confocal micrograph of phalloidin-labeled (green) isolated BBs shows that AM1-43 (red) evenly labels the BB membrane. Bars, 4 μm. (C) Increasing concentrations of BBs were treated with 2 mM ATP and the amount of membrane shed from each sample was measured using the microplate assay. ATP-induced membrane shedding scales with the BB concentration, demonstrating a linear response over two orders of magnitude. Points on this plot represent mean ± SD calculated from replicates at each BB concentration (for all values, SD is smaller than the point size).

Techniques Used: Labeling, Isolation, Concentration Assay

15) Product Images from "Cloning and characterization of prophenoloxidase A3 (proPOA3) from Culex pipiens pallens"

Article Title: Cloning and characterization of prophenoloxidase A3 (proPOA3) from Culex pipiens pallens

Journal: Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology

doi: 10.1016/j.cbpb.2012.04.008

Overexpressed proPOA3 leads to an increased susceptibility to deltamethrin in C6/36 cells. C6/36- proPOA3 and C6/36-control cells were treated with deltamethrin at the indicated concentrations, and viable cells were measured at 450 nm using a microplate
Figure Legend Snippet: Overexpressed proPOA3 leads to an increased susceptibility to deltamethrin in C6/36 cells. C6/36- proPOA3 and C6/36-control cells were treated with deltamethrin at the indicated concentrations, and viable cells were measured at 450 nm using a microplate

Techniques Used:

16) Product Images from "RAFT-based tri-component fluorescent glycopolymers: synthesis, characterization and application in lectin-mediated bacterial binding study"

Article Title: RAFT-based tri-component fluorescent glycopolymers: synthesis, characterization and application in lectin-mediated bacterial binding study

Journal: Glycoconjugate Journal

doi: 10.1007/s10719-013-9508-4

S. aureus ATCC 25923 showed binding specificity towards glycopolymers containing β-galactose as the pendant sugar. Different fluorescent glycopolymers (100 μg) were used in corresponding binding test, and 100 μL of the final bacteria suspensions in PBS were used to measure their fluorescence intensities on the microplate reader (λex/λem = 490/520 nm, slit width = 10 nm)
Figure Legend Snippet: S. aureus ATCC 25923 showed binding specificity towards glycopolymers containing β-galactose as the pendant sugar. Different fluorescent glycopolymers (100 μg) were used in corresponding binding test, and 100 μL of the final bacteria suspensions in PBS were used to measure their fluorescence intensities on the microplate reader (λex/λem = 490/520 nm, slit width = 10 nm)

Techniques Used: Binding Assay, Fluorescence

17) Product Images from "Luteoloside Inhibits Proliferation and Promotes Intrinsic and Extrinsic Pathway-Mediated Apoptosis Involving MAPK and mTOR Signaling Pathways in Human Cervical Cancer Cells"

Article Title: Luteoloside Inhibits Proliferation and Promotes Intrinsic and Extrinsic Pathway-Mediated Apoptosis Involving MAPK and mTOR Signaling Pathways in Human Cervical Cancer Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19061664

Effects of luteoloside on intracellular reactive oxygen species (ROS) levels in Hela cells. Cells were treated with various concentrations of luteoloside for the indicated time, and then incubated with 10 μM 2′,7′-dichlorodihydrofluorescein diacetate for 20 min. 2′,7′-dichlorodihydrofluorescein fluorescence was assayed on a microplate reader. ROS levels were given as the percentage of control without luteoloside treatment. The data are the mean ± SD of three independent experiments. *** p
Figure Legend Snippet: Effects of luteoloside on intracellular reactive oxygen species (ROS) levels in Hela cells. Cells were treated with various concentrations of luteoloside for the indicated time, and then incubated with 10 μM 2′,7′-dichlorodihydrofluorescein diacetate for 20 min. 2′,7′-dichlorodihydrofluorescein fluorescence was assayed on a microplate reader. ROS levels were given as the percentage of control without luteoloside treatment. The data are the mean ± SD of three independent experiments. *** p

Techniques Used: Incubation, Fluorescence

18) Product Images from "Luteoloside Inhibits Proliferation and Promotes Intrinsic and Extrinsic Pathway-Mediated Apoptosis Involving MAPK and mTOR Signaling Pathways in Human Cervical Cancer Cells"

Article Title: Luteoloside Inhibits Proliferation and Promotes Intrinsic and Extrinsic Pathway-Mediated Apoptosis Involving MAPK and mTOR Signaling Pathways in Human Cervical Cancer Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19061664

Effects of luteoloside on intracellular reactive oxygen species (ROS) levels in Hela cells. Cells were treated with various concentrations of luteoloside for the indicated time, and then incubated with 10 μM 2′,7′-dichlorodihydrofluorescein diacetate for 20 min. 2′,7′-dichlorodihydrofluorescein fluorescence was assayed on a microplate reader. ROS levels were given as the percentage of control without luteoloside treatment. The data are the mean ± SD of three independent experiments. *** p
Figure Legend Snippet: Effects of luteoloside on intracellular reactive oxygen species (ROS) levels in Hela cells. Cells were treated with various concentrations of luteoloside for the indicated time, and then incubated with 10 μM 2′,7′-dichlorodihydrofluorescein diacetate for 20 min. 2′,7′-dichlorodihydrofluorescein fluorescence was assayed on a microplate reader. ROS levels were given as the percentage of control without luteoloside treatment. The data are the mean ± SD of three independent experiments. *** p

Techniques Used: Incubation, Fluorescence

19) Product Images from "Isolation, Development, and Genomic Analysis of Bacillus megaterium SR7 for Growth and Metabolite Production Under Supercritical Carbon Dioxide"

Article Title: Isolation, Development, and Genomic Analysis of Bacillus megaterium SR7 for Growth and Metabolite Production Under Supercritical Carbon Dioxide

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.02152

Effect of mixing rates on SR7 growth in LB medium under 1 atm CO 2 for cultures inoculated from spores (A,B) or vegetative cells (C,D) . Growth was measured by CFUs/ml (A,C) and OD 600 (B,D) . Optical density was measured in 200 μl using a microplate reader and error bars represent the standard deviation of triplicate cultures. As expected, lag times for cultures inoculated with spores were longer than for those inoculated with vegetative cells due to time required for germination.
Figure Legend Snippet: Effect of mixing rates on SR7 growth in LB medium under 1 atm CO 2 for cultures inoculated from spores (A,B) or vegetative cells (C,D) . Growth was measured by CFUs/ml (A,C) and OD 600 (B,D) . Optical density was measured in 200 μl using a microplate reader and error bars represent the standard deviation of triplicate cultures. As expected, lag times for cultures inoculated with spores were longer than for those inoculated with vegetative cells due to time required for germination.

Techniques Used: Standard Deviation

SR7 growth under 1 atm CO 2 , 37°C, 250 rpm (OD 600 : red; CFU/ml: blue) (A) in LB medium (OD on linear scale), (B) in LB medium (OD on logarithmic scale), (C) in LB medium amended with glucose and/or trace metals, (D) in M9 salts + 50 mg/l YE + 4 g/l glucose as a function of trace metals concentration, (E) in M9+ minimal medium (% glucose consumption: green; OD on linear scale), and (F) in M9+ minimal medium (OD on logarithmic scale). OD 600 measured on 200 μl well microplate reader. Error bars represent standard deviation of triplicate cultures.
Figure Legend Snippet: SR7 growth under 1 atm CO 2 , 37°C, 250 rpm (OD 600 : red; CFU/ml: blue) (A) in LB medium (OD on linear scale), (B) in LB medium (OD on logarithmic scale), (C) in LB medium amended with glucose and/or trace metals, (D) in M9 salts + 50 mg/l YE + 4 g/l glucose as a function of trace metals concentration, (E) in M9+ minimal medium (% glucose consumption: green; OD on linear scale), and (F) in M9+ minimal medium (OD on logarithmic scale). OD 600 measured on 200 μl well microplate reader. Error bars represent standard deviation of triplicate cultures.

Techniques Used: Concentration Assay, Standard Deviation

20) Product Images from "Determination of Neuraminidase Kinetic Constants Using Whole Influenza Virus Preparations and Correction for Spectroscopic Interference by a Fluorogenic Substrate"

Article Title: Determination of Neuraminidase Kinetic Constants Using Whole Influenza Virus Preparations and Correction for Spectroscopic Interference by a Fluorogenic Substrate

Journal: PLoS ONE

doi: 10.1371/journal.pone.0071401

Standard curve of 4-methylumbelliferone (4-MU) fluorescence. Fluorescence intensity was measured using a Synergy 2 multimode microplate reader at excitation and emission wavelengths of 360 nm and 460 nm, respectively. Relative fluorescence units (RFU) obtained at low 4-MU concentrations (0–2.0 µM) are shown in the insert. Each data point represents the mean ± standard deviation (SD) of 10 independent measurements.
Figure Legend Snippet: Standard curve of 4-methylumbelliferone (4-MU) fluorescence. Fluorescence intensity was measured using a Synergy 2 multimode microplate reader at excitation and emission wavelengths of 360 nm and 460 nm, respectively. Relative fluorescence units (RFU) obtained at low 4-MU concentrations (0–2.0 µM) are shown in the insert. Each data point represents the mean ± standard deviation (SD) of 10 independent measurements.

Techniques Used: Fluorescence, Standard Deviation

Inner filter effect (IFE) from light absorption at the excitation and emission wavelengths of the 4-MU product. (A) Absorbance spectra of MUNANA and 4-MU at 0.01 mg/mL and 0.1 mg/mL concentrations in enzyme buffer. Optical density was measured using Synergy 2 multi-mode microplate reader in a UV-transparent 96-well plate. (B) Absorbance spectra of MUNANA and 4-MU at 0.1 mg/mL concentration. (C) 4-MU fluorescence measured in the presence of different concentrations of MUNANA (15–2000 µM) shows similar impact of MUNANA-associated spectroscopic interference across 4-MU concentrations of 10–80 µM.
Figure Legend Snippet: Inner filter effect (IFE) from light absorption at the excitation and emission wavelengths of the 4-MU product. (A) Absorbance spectra of MUNANA and 4-MU at 0.01 mg/mL and 0.1 mg/mL concentrations in enzyme buffer. Optical density was measured using Synergy 2 multi-mode microplate reader in a UV-transparent 96-well plate. (B) Absorbance spectra of MUNANA and 4-MU at 0.1 mg/mL concentration. (C) 4-MU fluorescence measured in the presence of different concentrations of MUNANA (15–2000 µM) shows similar impact of MUNANA-associated spectroscopic interference across 4-MU concentrations of 10–80 µM.

Techniques Used: Concentration Assay, Fluorescence

21) Product Images from "A novel chimeric lysin with robust antibacterial activity against planktonic and biofilm methicillin-resistant Staphylococcus aureus"

Article Title: A novel chimeric lysin with robust antibacterial activity against planktonic and biofilm methicillin-resistant Staphylococcus aureus

Journal: Scientific Reports

doi: 10.1038/srep40182

Characteristic of ClyF. ( A ) Time-killing curves of ClyF against S. aureus. S. aureus N315 were washed once with PBS and then treated with either 25 μg/ml ClyF, or equivalent volume of PBS buffer only, the changes of OD 600nm were monitored by a microplate reader at 37 °C for 15 min (left-Y axis, blank lines). Meanwhile, the viable cell number was calculated by plating onto TSB agar plates at various times (0, 2, 5, and 15 min, right-Y axis, blue lines). The effect of temperature ( B ), pH ( C ), EDTA (left Y-axis in ( D ), blank line) and NaCl (right Y-axis in ( D ), blue line) on the lytic activity of ClyF. The lytic activity of ClyF in each condition was compared and normalized as activity relative to the maximal activity. ( E – G ) Storage stability of ClyF. ClyF was stored at RT ( E ), 4 °C ( F ) and −80 °C ( G ) for different times (0–10 months). The residual activity against S. aureus N315 was determined by a microplate reader at 37 °C for 15 min, related to the activity of the fresh ClyF before storage. ( H ) Circular dichroism spectra of ClyF and Pc. ClyF (7.7 μM) and Pc (10.7 μM) in 4 mM Tris-HCl (pH 7.4) were scanned by a circular dichroism spectrometer from 190–260 nm at RT. ( I ) Thermal denaturation curves of ClyF and Pc. ClyF (38.5 μM) and Pc (21.4 μM) in 4 mM Tris-HCl (pH 7.4) were monitored by a circular dichroism spectrometer at 220 nm at temperatures from 25–90 °C. The Tm of each protein is estimated by the instrument’s software. Groups treated with PBS were used as controls, each assay was repeated for at least three times. Mean values are plotted, and error bars represent 1× standard deviation.
Figure Legend Snippet: Characteristic of ClyF. ( A ) Time-killing curves of ClyF against S. aureus. S. aureus N315 were washed once with PBS and then treated with either 25 μg/ml ClyF, or equivalent volume of PBS buffer only, the changes of OD 600nm were monitored by a microplate reader at 37 °C for 15 min (left-Y axis, blank lines). Meanwhile, the viable cell number was calculated by plating onto TSB agar plates at various times (0, 2, 5, and 15 min, right-Y axis, blue lines). The effect of temperature ( B ), pH ( C ), EDTA (left Y-axis in ( D ), blank line) and NaCl (right Y-axis in ( D ), blue line) on the lytic activity of ClyF. The lytic activity of ClyF in each condition was compared and normalized as activity relative to the maximal activity. ( E – G ) Storage stability of ClyF. ClyF was stored at RT ( E ), 4 °C ( F ) and −80 °C ( G ) for different times (0–10 months). The residual activity against S. aureus N315 was determined by a microplate reader at 37 °C for 15 min, related to the activity of the fresh ClyF before storage. ( H ) Circular dichroism spectra of ClyF and Pc. ClyF (7.7 μM) and Pc (10.7 μM) in 4 mM Tris-HCl (pH 7.4) were scanned by a circular dichroism spectrometer from 190–260 nm at RT. ( I ) Thermal denaturation curves of ClyF and Pc. ClyF (38.5 μM) and Pc (21.4 μM) in 4 mM Tris-HCl (pH 7.4) were monitored by a circular dichroism spectrometer at 220 nm at temperatures from 25–90 °C. The Tm of each protein is estimated by the instrument’s software. Groups treated with PBS were used as controls, each assay was repeated for at least three times. Mean values are plotted, and error bars represent 1× standard deviation.

Techniques Used: Activity Assay, Software, Standard Deviation

22) Product Images from "Impact of Hydroxychloroquine-Loaded Polyurethane Intravaginal Rings on Lactobacilli"

Article Title: Impact of Hydroxychloroquine-Loaded Polyurethane Intravaginal Rings on Lactobacilli

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.01819-15

Effects of HCQ treatment (24 h) on cell growth and viability. (A) Effects on Lactobacillus crispatus and Lactobacillus jensenii . Bacterial growth was measured at 580 nm using a microplate reader. Lactobacilli treated with drug-free medium were used as
Figure Legend Snippet: Effects of HCQ treatment (24 h) on cell growth and viability. (A) Effects on Lactobacillus crispatus and Lactobacillus jensenii . Bacterial growth was measured at 580 nm using a microplate reader. Lactobacilli treated with drug-free medium were used as

Techniques Used:

23) Product Images from "Induction of enhanced immunogenic cell death through ultrasound-controlled release of doxorubicin by liposome-microbubble complexes"

Article Title: Induction of enhanced immunogenic cell death through ultrasound-controlled release of doxorubicin by liposome-microbubble complexes

Journal: Oncoimmunology

doi: 10.1080/2162402X.2018.1446720

Increased intracellular uptake and tissue accumulation of Dox by MbDox+US treatment. A. LL/2 and CT26 cells were treated with indicated formulations at 4 h after treatment. Images of intracellular Dox distribution were collected by confocal microscopy. Dox is indicated by red fluorescence and the nucleus is stained with DAPI (blue, scale bar = 10 μm). B. The intracellular concentration of Dox in LL/2 and CT26 cells after treatment with the indicated formulations was quantified by a microplate reader. C. Tumor-bearing mice were treated with indicated formulations, and tumor tissues and major organs were collected to analyze the Dox concentration by HPLC. Data are expressed as mean ± SEM, * indicates P
Figure Legend Snippet: Increased intracellular uptake and tissue accumulation of Dox by MbDox+US treatment. A. LL/2 and CT26 cells were treated with indicated formulations at 4 h after treatment. Images of intracellular Dox distribution were collected by confocal microscopy. Dox is indicated by red fluorescence and the nucleus is stained with DAPI (blue, scale bar = 10 μm). B. The intracellular concentration of Dox in LL/2 and CT26 cells after treatment with the indicated formulations was quantified by a microplate reader. C. Tumor-bearing mice were treated with indicated formulations, and tumor tissues and major organs were collected to analyze the Dox concentration by HPLC. Data are expressed as mean ± SEM, * indicates P

Techniques Used: Confocal Microscopy, Fluorescence, Staining, Concentration Assay, Mouse Assay, High Performance Liquid Chromatography

24) Product Images from "Metformin and gefitinib cooperate to inhibit bladder cancer growth via both AMPK and EGFR pathways joining at Akt and Erk"

Article Title: Metformin and gefitinib cooperate to inhibit bladder cancer growth via both AMPK and EGFR pathways joining at Akt and Erk

Journal: Scientific Reports

doi: 10.1038/srep28611

Evaluation of colony suppression of Gefitinib alone or combined with metformin on 3 bladder cancer cell lines. ( A–C ) Clonogenic assay was assessed after 7 day Gefitinib treatment alone or combined with metformin and wells were stained with crystal violet at the end of the experiment. ( A ) Clonogenic assay in MB49 was conducted with the treatment of 2 mM metformin, 0.4 μM Gefitinib and their combination. Above: The full view of wells were taken through stereomicroscope and images were taken through an inverted microscope with ×10 magnification. Below: the quantification of colony was determined by microplate area scan at OD 550 nm, Results are presented as the median of 5 independent experiments (*P
Figure Legend Snippet: Evaluation of colony suppression of Gefitinib alone or combined with metformin on 3 bladder cancer cell lines. ( A–C ) Clonogenic assay was assessed after 7 day Gefitinib treatment alone or combined with metformin and wells were stained with crystal violet at the end of the experiment. ( A ) Clonogenic assay in MB49 was conducted with the treatment of 2 mM metformin, 0.4 μM Gefitinib and their combination. Above: The full view of wells were taken through stereomicroscope and images were taken through an inverted microscope with ×10 magnification. Below: the quantification of colony was determined by microplate area scan at OD 550 nm, Results are presented as the median of 5 independent experiments (*P

Techniques Used: Clonogenic Assay, Staining, Inverted Microscopy

25) Product Images from "Effects of a Chimeric Lysin against Planktonic and Sessile Enterococcus faecalis Hint at Potential Application in Endodontic Therapy"

Article Title: Effects of a Chimeric Lysin against Planktonic and Sessile Enterococcus faecalis Hint at Potential Application in Endodontic Therapy

Journal: Viruses

doi: 10.3390/v10060290

High activity of ClyR against E. faecalis biofilm. ( a ) Crystal violet staining analysis of E. faecalis biofilms. E. faecalis ATCC 51299 cells are cultured in BHI supplemented with 3% glucose (BHIG) for 24 h to allow biofilm formation, after washing with PBS, biofilms are treated with various concentrations of ClyR (0, 50, 100, and 200 μg/mL) at 37 °C for 1 h, residual biofilms are stained with 0.1% crystal violet and detected at OD 595 by a microplate reader after resolved by ethanol. ( b ) Survival rate of E. faecalis biofilms after treatment with ClyR. 24 h-aged E. faecalis ATCC 51299 biofilms are treated with various concentrations of ClyR (0, 50, 100, and 200 μg/mL) at 37 °C for 1 h, the viable cells after each treatment is determined and compared with that of PBS treated controls by Two-tailed Student’s t test. Data is shown as mean ± standard deviation, and ** p
Figure Legend Snippet: High activity of ClyR against E. faecalis biofilm. ( a ) Crystal violet staining analysis of E. faecalis biofilms. E. faecalis ATCC 51299 cells are cultured in BHI supplemented with 3% glucose (BHIG) for 24 h to allow biofilm formation, after washing with PBS, biofilms are treated with various concentrations of ClyR (0, 50, 100, and 200 μg/mL) at 37 °C for 1 h, residual biofilms are stained with 0.1% crystal violet and detected at OD 595 by a microplate reader after resolved by ethanol. ( b ) Survival rate of E. faecalis biofilms after treatment with ClyR. 24 h-aged E. faecalis ATCC 51299 biofilms are treated with various concentrations of ClyR (0, 50, 100, and 200 μg/mL) at 37 °C for 1 h, the viable cells after each treatment is determined and compared with that of PBS treated controls by Two-tailed Student’s t test. Data is shown as mean ± standard deviation, and ** p

Techniques Used: Activity Assay, Staining, Cell Culture, Two Tailed Test, Standard Deviation

Robust and rapid bactericidal activity of ClyR against planktonic E. faecalis . ( a ) Lysis curves of ClyR against E. faecalis ATCC 51299. Bacterial cells are washed with phosphate-buffered saline (PBS) and mixed with various concentrations of ClyR (0, 12.5, 25, and 50 μg/mL), the decrease of OD 600 is monitored by a microplate reader for 30 min at 37 °C. ( b ) Dose-dependent killing efficacy of ClyR against E. faecalis ATCC51299. Bacterial cells are washed with PBS and treated with various concentrations of ClyR (0, 0.5, 2, 5, and 25 μg/mL) for 40 min at 37 °C, the viable cell number after each treatment is determined by plating on brain heart infusion (BHI) agar. ( c ) Time-dependent killing efficacy of ClyR against E. faecalis ATCC51299. Bacterial cells are washed with PBS and treated with 50 μg/mL of ClyR at 37 °C for different times (0, 0.5, 1, 5, and 15 min), the viable cell number after each treatment is determined by plating on BHI agar. ( d ) Susceptibility of multiple E. faecalis strains to ClyR. The result of each treatment is compared with that of the PBS-treated controls by Two-tailed Student’s t test. Data is shown as mean ± standard deviation, and * p
Figure Legend Snippet: Robust and rapid bactericidal activity of ClyR against planktonic E. faecalis . ( a ) Lysis curves of ClyR against E. faecalis ATCC 51299. Bacterial cells are washed with phosphate-buffered saline (PBS) and mixed with various concentrations of ClyR (0, 12.5, 25, and 50 μg/mL), the decrease of OD 600 is monitored by a microplate reader for 30 min at 37 °C. ( b ) Dose-dependent killing efficacy of ClyR against E. faecalis ATCC51299. Bacterial cells are washed with PBS and treated with various concentrations of ClyR (0, 0.5, 2, 5, and 25 μg/mL) for 40 min at 37 °C, the viable cell number after each treatment is determined by plating on brain heart infusion (BHI) agar. ( c ) Time-dependent killing efficacy of ClyR against E. faecalis ATCC51299. Bacterial cells are washed with PBS and treated with 50 μg/mL of ClyR at 37 °C for different times (0, 0.5, 1, 5, and 15 min), the viable cell number after each treatment is determined by plating on BHI agar. ( d ) Susceptibility of multiple E. faecalis strains to ClyR. The result of each treatment is compared with that of the PBS-treated controls by Two-tailed Student’s t test. Data is shown as mean ± standard deviation, and * p

Techniques Used: Activity Assay, Lysis, Two Tailed Test, Standard Deviation

26) Product Images from "Porous Electrospun Fibers with Self-Sealing Functionality: An Enabling Strategy for Trapping Biomacromolecules"

Article Title: Porous Electrospun Fibers with Self-Sealing Functionality: An Enabling Strategy for Trapping Biomacromolecules

Journal: Small (Weinheim an der Bergstrasse, Germany)

doi: 10.1002/smll.201701949

Characterizations of electrospun fibers with different CNT amounts. A) Gross images and B) representative SEM micrographs of PLA/CNT composite fibers. C) TGA thermograms under a nitrogen atmosphere and D) absorbance at 650 nm measured by microplate reader of electrospun PLA/CNT fibers with different CNT concentrations. Results are presented as means ± standard deviations ( n = 3; * p
Figure Legend Snippet: Characterizations of electrospun fibers with different CNT amounts. A) Gross images and B) representative SEM micrographs of PLA/CNT composite fibers. C) TGA thermograms under a nitrogen atmosphere and D) absorbance at 650 nm measured by microplate reader of electrospun PLA/CNT fibers with different CNT concentrations. Results are presented as means ± standard deviations ( n = 3; * p

Techniques Used: Proximity Ligation Assay

27) Product Images from "Enhanced Antibacterial Activity of Acinetobacter baumannii Bacteriophage ØABP-01 Endolysin (LysABP-01) in Combination with Colistin"

Article Title: Enhanced Antibacterial Activity of Acinetobacter baumannii Bacteriophage ØABP-01 Endolysin (LysABP-01) in Combination with Colistin

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2016.01402

Antibacterial activity of LysABP-01. (A) Bacterial growth inhibition activity of varying concentrations of LysABP-01. The bar graph depicts the percent growth inhibition and represents the mean percentage ± SD of triplicate experiments. (B) The representative microplate image of TTC staining.
Figure Legend Snippet: Antibacterial activity of LysABP-01. (A) Bacterial growth inhibition activity of varying concentrations of LysABP-01. The bar graph depicts the percent growth inhibition and represents the mean percentage ± SD of triplicate experiments. (B) The representative microplate image of TTC staining.

Techniques Used: Activity Assay, Inhibition, Staining

28) Product Images from "Conversion of Cytochrome P450 2D6 of Human Into a FRET-Based Tool for Real-Time Monitoring of Ajmalicine in Living Cells"

Article Title: Conversion of Cytochrome P450 2D6 of Human Into a FRET-Based Tool for Real-Time Monitoring of Ajmalicine in Living Cells

Journal: Frontiers in Bioengineering and Biotechnology

doi: 10.3389/fbioe.2019.00375

Fluorescence emission intensity recorded in a microplate reader in the absence and presence of 1 mM ajmalicine. Advancement in the fluorescence intensity of Venus is presented.
Figure Legend Snippet: Fluorescence emission intensity recorded in a microplate reader in the absence and presence of 1 mM ajmalicine. Advancement in the fluorescence intensity of Venus is presented.

Techniques Used: Fluorescence

29) Product Images from "Modulation of Radiation Injury Response in Retinal Endothelial Cells by Quinic Acid Derivative KZ-41 Involves p38 MAPK"

Article Title: Modulation of Radiation Injury Response in Retinal Endothelial Cells by Quinic Acid Derivative KZ-41 Involves p38 MAPK

Journal: PLoS ONE

doi: 10.1371/journal.pone.0100210

Radiation-induced REC-U937 adhesion. Panel A) Static-adhesion: Irradiated human RECs were incubated for 24 hours in culture medium containing vehicle (PBS) or KZ-41 (10 µM). Calcein-AM loaded U937 cells were co-cultured with RECs (n = 8/group) for 30 minutes and non-adherent cells were washed from wells; attached U937 cells were quantified with a fluorescence microplate reader (excitation/emission wavelengths of 485/535 ηm). Data demonstrate that KZ-41 inhibits IR-induced leukocyte attachment to RECs (*, **P
Figure Legend Snippet: Radiation-induced REC-U937 adhesion. Panel A) Static-adhesion: Irradiated human RECs were incubated for 24 hours in culture medium containing vehicle (PBS) or KZ-41 (10 µM). Calcein-AM loaded U937 cells were co-cultured with RECs (n = 8/group) for 30 minutes and non-adherent cells were washed from wells; attached U937 cells were quantified with a fluorescence microplate reader (excitation/emission wavelengths of 485/535 ηm). Data demonstrate that KZ-41 inhibits IR-induced leukocyte attachment to RECs (*, **P

Techniques Used: Irradiation, Incubation, Cell Culture, Fluorescence

30) Product Images from "Bioinspired oral delivery of gut microbiota by self-coating with biofilms"

Article Title: Bioinspired oral delivery of gut microbiota by self-coating with biofilms

Journal: Science Advances

doi: 10.1126/sciadv.abb1952

Mucoadhesion of self-coated bacteria in mouse intestines. ( A to C ) Numbers of BS, FCBS, and BCBS that adhered to the (A) small intestine, (B) large intestine, and (C) cecum at 4, 48, and 120 hours after administration, respectively. Each mouse was fed with 1 × 10 7 CFUs of bacteria by oral gavage and then sacrificed at the indicated time points. Bacteria were quantified by plate counting. Error bars represent SD ( n = 5). ( D ) Representative microscopic images of Gram staining of the intestinal tissues harvested from mice orally delivered with 1 × 10 7 CFUs of bacteria at 24 hours after administration. Scale bars, 100 μm. ( E ) Growth curves of bacteria cultured in SIF at 37°C, and OD 600 was recorded at 30-min intervals using a microplate reader. ( F ) Bacterial viability in SIF (pH 6.8). Error bars represent SD ( n = 3). ( G ) Typical TEM images of bacteria after culture in SIF at 37°C for the indicated time points. Scale bars, 1 μm. * P
Figure Legend Snippet: Mucoadhesion of self-coated bacteria in mouse intestines. ( A to C ) Numbers of BS, FCBS, and BCBS that adhered to the (A) small intestine, (B) large intestine, and (C) cecum at 4, 48, and 120 hours after administration, respectively. Each mouse was fed with 1 × 10 7 CFUs of bacteria by oral gavage and then sacrificed at the indicated time points. Bacteria were quantified by plate counting. Error bars represent SD ( n = 5). ( D ) Representative microscopic images of Gram staining of the intestinal tissues harvested from mice orally delivered with 1 × 10 7 CFUs of bacteria at 24 hours after administration. Scale bars, 100 μm. ( E ) Growth curves of bacteria cultured in SIF at 37°C, and OD 600 was recorded at 30-min intervals using a microplate reader. ( F ) Bacterial viability in SIF (pH 6.8). Error bars represent SD ( n = 3). ( G ) Typical TEM images of bacteria after culture in SIF at 37°C for the indicated time points. Scale bars, 1 μm. * P

Techniques Used: Staining, Mouse Assay, Cell Culture, Transmission Electron Microscopy

31) Product Images from "Genetic variation in the MacAB-TolC efflux pump influences pathogenesis of invasive Salmonella isolates from Africa"

Article Title: Genetic variation in the MacAB-TolC efflux pump influences pathogenesis of invasive Salmonella isolates from Africa

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1008763

S . Typhimurium ST19 macAB provides superior resistance to the antimicrobial peptide C18G. 4/74 phoP acrAB macAB null mutants with low-copy pACYC177 plasmids constitutively-expressing macAB variants (pJH14-17, see S3 Table ) were grown in N minimal medium, pH 7.4 and 1mM MgCl 2 . Overnight stationary phase cells were washed and normalized to OD600 = 1 before 1:200 final dilution into fresh N minimal medium with 2μg/mL C18G (A) or 1μg/mL C18G (B). OD600 was monitored over time using a BioTek Synergy HTX plate reader. Growth curves presented here are from one experimental run and representative of three independent experiments. Plotted data points are the geometric mean of quadruplicate or triplicate microplate wells. Lag time (inset table) was determined as time to reach OD600 = 0.15.
Figure Legend Snippet: S . Typhimurium ST19 macAB provides superior resistance to the antimicrobial peptide C18G. 4/74 phoP acrAB macAB null mutants with low-copy pACYC177 plasmids constitutively-expressing macAB variants (pJH14-17, see S3 Table ) were grown in N minimal medium, pH 7.4 and 1mM MgCl 2 . Overnight stationary phase cells were washed and normalized to OD600 = 1 before 1:200 final dilution into fresh N minimal medium with 2μg/mL C18G (A) or 1μg/mL C18G (B). OD600 was monitored over time using a BioTek Synergy HTX plate reader. Growth curves presented here are from one experimental run and representative of three independent experiments. Plotted data points are the geometric mean of quadruplicate or triplicate microplate wells. Lag time (inset table) was determined as time to reach OD600 = 0.15.

Techniques Used: Expressing

32) Product Images from "Protective Effect of Simplicillium sp. Ethyl Acetate Extract against High Glucose-Induced Oxidative Stress in HUVECs"

Article Title: Protective Effect of Simplicillium sp. Ethyl Acetate Extract against High Glucose-Induced Oxidative Stress in HUVECs

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2020/5172765

Effect of CH on intracellular reactive oxygen species (ROS) production in HG-induced HUVECs. (a) Intracellular ROS levels in HUVECs were assessed DCFH-DA probe after treated with high glucose (HG, 35 mM) with or without CH for 24 h. HUVECs were loaded with DCFH-DA in the dark at 37°C for 40 min and then pictures were collected with a fluorescence microscope (magnification 200×). (b) Intracellular fluorescence was measured using a fluorescence microplate at excitation and emission wavelengths of 488 and 525 nm. Results were represented as the mean ± SEM ( n = 3). N.S P > 0.05 vs. control group; ## P
Figure Legend Snippet: Effect of CH on intracellular reactive oxygen species (ROS) production in HG-induced HUVECs. (a) Intracellular ROS levels in HUVECs were assessed DCFH-DA probe after treated with high glucose (HG, 35 mM) with or without CH for 24 h. HUVECs were loaded with DCFH-DA in the dark at 37°C for 40 min and then pictures were collected with a fluorescence microscope (magnification 200×). (b) Intracellular fluorescence was measured using a fluorescence microplate at excitation and emission wavelengths of 488 and 525 nm. Results were represented as the mean ± SEM ( n = 3). N.S P > 0.05 vs. control group; ## P

Techniques Used: Fluorescence, Microscopy

33) Product Images from "Syntrophic co-culture amplification of production phenotype for high-throughput screening of microbial strain libraries"

Article Title: Syntrophic co-culture amplification of production phenotype for high-throughput screening of microbial strain libraries

Journal: bioRxiv

doi: 10.1101/518639

Co-culture growth characteristics correlate with monoculture production performance. (A) 2-KIV and isobutanol production pathway. Genes that are overexpressed in the production strains are boxed in the color corresponding to the strain in the legend in (C). (B) Monoculture 2-KIV production characteristics of three different secretor strains on 20 g/L glucose. IPTG was added after 8 h and 2-KIV was measured 60 h after induction. Error bars represent the standard deviation of three biological replicates. (C) Monoculture isobutanol production characteristics of these three strains when carrying pSA65 P L lacO 1 :: kivd–adhA ). Isobutanol, glucose and cell growth profiles are shown. Error bars represent the standard deviation of four biological replicates. Growth profiles of co-cultures of the same three secretor strains (not carrying pSA65) with sensor strain K12 Δ ilvD::kan in a microplate with (D) and without (E) IPTG-induced expression of the alsS/ilvCD genes. Three replicate wells are shown for each strain pair, plotted in the same color. (F) Co-culture growth rates. Error bars represent the standard deviations between at least 10 biological replicates, combined from two separate experiments. (G) Population composition of microplate cultures in early stationary phase. Error bars represent standard deviations from two biological replicates with two technical replicates each.
Figure Legend Snippet: Co-culture growth characteristics correlate with monoculture production performance. (A) 2-KIV and isobutanol production pathway. Genes that are overexpressed in the production strains are boxed in the color corresponding to the strain in the legend in (C). (B) Monoculture 2-KIV production characteristics of three different secretor strains on 20 g/L glucose. IPTG was added after 8 h and 2-KIV was measured 60 h after induction. Error bars represent the standard deviation of three biological replicates. (C) Monoculture isobutanol production characteristics of these three strains when carrying pSA65 P L lacO 1 :: kivd–adhA ). Isobutanol, glucose and cell growth profiles are shown. Error bars represent the standard deviation of four biological replicates. Growth profiles of co-cultures of the same three secretor strains (not carrying pSA65) with sensor strain K12 Δ ilvD::kan in a microplate with (D) and without (E) IPTG-induced expression of the alsS/ilvCD genes. Three replicate wells are shown for each strain pair, plotted in the same color. (F) Co-culture growth rates. Error bars represent the standard deviations between at least 10 biological replicates, combined from two separate experiments. (G) Population composition of microplate cultures in early stationary phase. Error bars represent standard deviations from two biological replicates with two technical replicates each.

Techniques Used: Co-Culture Assay, Standard Deviation, Expressing

34) Product Images from "A novel calcium-concentrating compartment drives biofilm formation and persistent infections"

Article Title: A novel calcium-concentrating compartment drives biofilm formation and persistent infections

Journal: bioRxiv

doi: 10.1101/2020.01.08.898569

A. Transcription profile of B. subtilis NCIB 3610 biofilm colonies grown for 1, 3 and 6 days on a biofilm-inducing B4-Ca agar, with and without added excess calcium. A heatmap with all differentially expressed genes (n = 876), scaled to the mean expression level of each gene. The map is ordered by age of biofilm colonies, as shown by the left colored bar, where green are genes that are differentially expressed in days 1-3, and day 6; purple genes that are differentially expressed in days 1-3 and yellow genes that are differentially expressed in day 6. B. The top 20 conditions from the study of Nicolas et al (9), showing the highest similarity to the transcriptome of 6-day old biofilm colonies grown without or with calcium (top panel and bottom panel, respectively). C-D. Volcano-plot depicting calcium-dependent changes in expression of genes. Genes belonging to the indicated functional categories, as determined by DAVID analysis, are highlighted. E. Calcium-dependent change in gene expression of genes highlighted in D, expressed as log 2 fold change. F. Top-down images of biofilm colonies of wild-type B. subtilis strain and the indicated mutant derivatives, grown on solid biofilm-inducing medium B4, with added excess calcium, for 3 days at 30°C. G. Top-down images of biofilm colonies of wild-type B. subtilis grown for 3 days at 30°C on solid B4 biofilm-inducing medium, with added excess calcium, and supplemented with indicated inhibitors (0.1 mg/ml SMV, 2.5 mg/ml AHA). H. Planktonic growth assays. Wild type and the indicated mutant derivatives were grown at 30°C with shaking in liquid B4 medium supplemented with calcium, and growth was monitored by measuring OD 600 in a microplate reader every 30 min. Results are averages of nine wells, bars represent standard deviations. A representative experiment out of at least 3 independent experiments is shown.
Figure Legend Snippet: A. Transcription profile of B. subtilis NCIB 3610 biofilm colonies grown for 1, 3 and 6 days on a biofilm-inducing B4-Ca agar, with and without added excess calcium. A heatmap with all differentially expressed genes (n = 876), scaled to the mean expression level of each gene. The map is ordered by age of biofilm colonies, as shown by the left colored bar, where green are genes that are differentially expressed in days 1-3, and day 6; purple genes that are differentially expressed in days 1-3 and yellow genes that are differentially expressed in day 6. B. The top 20 conditions from the study of Nicolas et al (9), showing the highest similarity to the transcriptome of 6-day old biofilm colonies grown without or with calcium (top panel and bottom panel, respectively). C-D. Volcano-plot depicting calcium-dependent changes in expression of genes. Genes belonging to the indicated functional categories, as determined by DAVID analysis, are highlighted. E. Calcium-dependent change in gene expression of genes highlighted in D, expressed as log 2 fold change. F. Top-down images of biofilm colonies of wild-type B. subtilis strain and the indicated mutant derivatives, grown on solid biofilm-inducing medium B4, with added excess calcium, for 3 days at 30°C. G. Top-down images of biofilm colonies of wild-type B. subtilis grown for 3 days at 30°C on solid B4 biofilm-inducing medium, with added excess calcium, and supplemented with indicated inhibitors (0.1 mg/ml SMV, 2.5 mg/ml AHA). H. Planktonic growth assays. Wild type and the indicated mutant derivatives were grown at 30°C with shaking in liquid B4 medium supplemented with calcium, and growth was monitored by measuring OD 600 in a microplate reader every 30 min. Results are averages of nine wells, bars represent standard deviations. A representative experiment out of at least 3 independent experiments is shown.

Techniques Used: Expressing, Functional Assay, Mutagenesis

35) Product Images from "Ranking of Major Classes of Antibiotics for Activity against Stationary Phase Pseudomonas aeruginosa and Identification of Clinafloxacin + Cefuroxime + Gentamicin Drug Combination that Eradicates Persistent P. aeruginosa Infection in a Murine Cystic Fibrosis Model"

Article Title: Ranking of Major Classes of Antibiotics for Activity against Stationary Phase Pseudomonas aeruginosa and Identification of Clinafloxacin + Cefuroxime + Gentamicin Drug Combination that Eradicates Persistent P. aeruginosa Infection in a Murine Cystic Fibrosis Model

Journal: bioRxiv

doi: 10.1101/686105

SYBR Green/PI staining can assess the viability of P. aeruginosa (A). A standard curve revealed linear relationship between the Green/Red fluorescence ratios from SYBR Green/PI viability assay and the percentage of live P. aeruginosa PAO-1 cells. The dead organisms were prepared with 70 % isopropyl alcohol and different proportions of live and dead cells were mixed and stained with SYBR Green I/PI and the green/red fluorescence ratios were measured by a microplate reader. (B) Representative microscope images of 0%, 50% and 100% of live P. aeruginosa cells using SYBR Green I/PI stain (200 X magnification). Green cells represent live cells and Red cells represent dead cells.
Figure Legend Snippet: SYBR Green/PI staining can assess the viability of P. aeruginosa (A). A standard curve revealed linear relationship between the Green/Red fluorescence ratios from SYBR Green/PI viability assay and the percentage of live P. aeruginosa PAO-1 cells. The dead organisms were prepared with 70 % isopropyl alcohol and different proportions of live and dead cells were mixed and stained with SYBR Green I/PI and the green/red fluorescence ratios were measured by a microplate reader. (B) Representative microscope images of 0%, 50% and 100% of live P. aeruginosa cells using SYBR Green I/PI stain (200 X magnification). Green cells represent live cells and Red cells represent dead cells.

Techniques Used: SYBR Green Assay, Staining, Fluorescence, Viability Assay, Microscopy

36) Product Images from "Folic acid protects against arsenic-mediated embryo toxicity by up -regulating the expression of Dvr1"

Article Title: Folic acid protects against arsenic-mediated embryo toxicity by up -regulating the expression of Dvr1

Journal: Scientific Reports

doi: 10.1038/srep16093

Folic acid reduces ROS levels and the expression of p66Shc in arsenite-exposed HEK293ET cells. ( A ) and ( B ) Levels of ROS were assessed by fluorescence microscope ( A ) and microplate reader ( B ) in HEK293ET cells treated with arsenite, folic acid and their combination. ( C ) Expression of p66Shc was assessed by Western blot in HEK293ET cells treated with different concentrations of arsenite. Quantification of p66Shc expression by densitometry. ( D ) Expression of p66Shc was assessed by Western blot in HEK293ET cells treated with arsenite, folic acid and their combination. Quantification of p66Shc expression by densitometry. * p
Figure Legend Snippet: Folic acid reduces ROS levels and the expression of p66Shc in arsenite-exposed HEK293ET cells. ( A ) and ( B ) Levels of ROS were assessed by fluorescence microscope ( A ) and microplate reader ( B ) in HEK293ET cells treated with arsenite, folic acid and their combination. ( C ) Expression of p66Shc was assessed by Western blot in HEK293ET cells treated with different concentrations of arsenite. Quantification of p66Shc expression by densitometry. ( D ) Expression of p66Shc was assessed by Western blot in HEK293ET cells treated with arsenite, folic acid and their combination. Quantification of p66Shc expression by densitometry. * p

Techniques Used: Expressing, Fluorescence, Microscopy, Western Blot

37) Product Images from "Regulation of differential proton-coupled folate transporter gene expression in human tumors: transactivation by KLF15 with NRF-1 and the role of Sp1"

Article Title: Regulation of differential proton-coupled folate transporter gene expression in human tumors: transactivation by KLF15 with NRF-1 and the role of Sp1

Journal: The Biochemical journal

doi: 10.1042/BCJ20180394

Functional analysis of hPCFT core promoter elements and transcription factors in Drosophila SL2 cells. Drosophila SL2 cells were co-transfected with hPCFT core promoter-luciferase reporter gene constructs −50/+96 and pPacO, or Sp1, USp3 (Sp3 longer form), Sp3 (Sp3 shorter form), NRF-1, or KLF15 cDNA constructs in pPacO using FuGENE 6 transfection reagent. Cells were harvested after 48 h for luciferase assays using a single-luciferase assay system with a microplate reader. Luciferase activities were normalized to cell proteins. ( A ) Effects of assorted transcription factors (Sp1, Sp3, USp3, NRF-1, and KLF15; added singly or in combination) on hPCFT core promoter activity. Statistics were performed within each transactivation group by Sp1, NRF-1, KLF15, or NRF-1 + KLF15, upon addition of various Sp factors. ( B ) Transactivation of WT hPCFT core promoter (−50/+96) and hPCFT core promoter with the GC-Box consensus sequence mutation (−50/+96/GC-Boxm), upon transfecting with Sp1, NRF-1, or KLF15 singly or in combination. For both panels A and B, results are presented as mean values ± standard errors from 3 to 11 different experiments. For both panels: **** p
Figure Legend Snippet: Functional analysis of hPCFT core promoter elements and transcription factors in Drosophila SL2 cells. Drosophila SL2 cells were co-transfected with hPCFT core promoter-luciferase reporter gene constructs −50/+96 and pPacO, or Sp1, USp3 (Sp3 longer form), Sp3 (Sp3 shorter form), NRF-1, or KLF15 cDNA constructs in pPacO using FuGENE 6 transfection reagent. Cells were harvested after 48 h for luciferase assays using a single-luciferase assay system with a microplate reader. Luciferase activities were normalized to cell proteins. ( A ) Effects of assorted transcription factors (Sp1, Sp3, USp3, NRF-1, and KLF15; added singly or in combination) on hPCFT core promoter activity. Statistics were performed within each transactivation group by Sp1, NRF-1, KLF15, or NRF-1 + KLF15, upon addition of various Sp factors. ( B ) Transactivation of WT hPCFT core promoter (−50/+96) and hPCFT core promoter with the GC-Box consensus sequence mutation (−50/+96/GC-Boxm), upon transfecting with Sp1, NRF-1, or KLF15 singly or in combination. For both panels A and B, results are presented as mean values ± standard errors from 3 to 11 different experiments. For both panels: **** p

Techniques Used: Functional Assay, Transfection, Luciferase, Construct, Activity Assay, Sequencing, Mutagenesis

38) Product Images from "Targeting Methicillin-Resistant Staphylococcus aureus with Short Salt-Resistant Synthetic Peptides"

Article Title: Targeting Methicillin-Resistant Staphylococcus aureus with Short Salt-Resistant Synthetic Peptides

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.02578-14

Killing kinetics of MRSA USA300 exposed to 4× MIC of RRIKA, RR, nisin, vancomycin, and sterile water (control) as observed by measuring the OD 595 by microplate reader over time. Results are representative of two separate experiments, each done in triplicate. Error bars represent standard deviation values (data without error bars indicate that the SD is too small to be seen).
Figure Legend Snippet: Killing kinetics of MRSA USA300 exposed to 4× MIC of RRIKA, RR, nisin, vancomycin, and sterile water (control) as observed by measuring the OD 595 by microplate reader over time. Results are representative of two separate experiments, each done in triplicate. Error bars represent standard deviation values (data without error bars indicate that the SD is too small to be seen).

Techniques Used: Standard Deviation

39) Product Images from "Oxidative Stress Mediates Microcystin-LR-Induced Endoplasmic Reticulum Stress and Autophagy in KK-1 Cells and C57BL/6 Mice Ovaries"

Article Title: Oxidative Stress Mediates Microcystin-LR-Induced Endoplasmic Reticulum Stress and Autophagy in KK-1 Cells and C57BL/6 Mice Ovaries

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2018.01058

The effect of N -acetyl-l-cysteine (NAC) and Microcystin-LR (MC-LR) on redox-related indicators in KK-1 cells and C57BL/6 mice ovarian tissues. (A) The activity of superoxide dismutase (SOD) was measured in KK-1 cells. (B) glutathione/oxidized glutathione (GSH/GSSG) content was determined via the colorimetric method in KK-1 cells. (C) The levels of reactive oxygen species (ROS) in mice ovarian tissues were detected using an automated microplate reader via monitoring DCF fluorescence intensity. (D) Malondialdehyde (MDA) was measured using a colorimetric assay kit in mice ovarian tissues. (E) WST-8 method was used to measure the activity of SOD in mice ovarian tissues. (F) GSH/GSSG content was determined in mice ovarian tissues. Data were expressed as mean ± standard deviation (SD, n = 3), ∗ P
Figure Legend Snippet: The effect of N -acetyl-l-cysteine (NAC) and Microcystin-LR (MC-LR) on redox-related indicators in KK-1 cells and C57BL/6 mice ovarian tissues. (A) The activity of superoxide dismutase (SOD) was measured in KK-1 cells. (B) glutathione/oxidized glutathione (GSH/GSSG) content was determined via the colorimetric method in KK-1 cells. (C) The levels of reactive oxygen species (ROS) in mice ovarian tissues were detected using an automated microplate reader via monitoring DCF fluorescence intensity. (D) Malondialdehyde (MDA) was measured using a colorimetric assay kit in mice ovarian tissues. (E) WST-8 method was used to measure the activity of SOD in mice ovarian tissues. (F) GSH/GSSG content was determined in mice ovarian tissues. Data were expressed as mean ± standard deviation (SD, n = 3), ∗ P

Techniques Used: Mouse Assay, Activity Assay, Fluorescence, Multiple Displacement Amplification, Colorimetric Assay, Standard Deviation

40) Product Images from "Salidroside Suppresses HUVECs Cell Injury Induced by Oxidative Stress through Activating the Nrf2 Signaling Pathway"

Article Title: Salidroside Suppresses HUVECs Cell Injury Induced by Oxidative Stress through Activating the Nrf2 Signaling Pathway

Journal: Molecules

doi: 10.3390/molecules21081033

SAL reduces H 2 O 2 -induced ROS production and MDA formation and increases the activities of antioxidant enzymes in HUVECs. ( a ) HUVECs were treated with various concentrations of H 2 O 2 (0–500 μM) for 4 h, and cell viability was determined by MTT assay. ( b – e ) HUVECs were incubated without any intervention (control), SAL (0.1, 1, 10 μM) for 24 h, or 300 μM H 2 O 2 for 4 h, or 0.1, 1, 10 μM SAL for 24 h followed by 300 μM H 2 O 2 for an additional 4 h. ( b ) Intercellular ROS levels: Intercellular ROS levels were determined by DCFH-DA fluorescence. After various treatment, the cells were stained with 10 μM DCFH-DA in serum free medium for 30 min in the dark. Fluorescence intensity was recorded using a Synergy TM 4 Multi-Mode Microplate Reader at an excitation wavelength of 488 nm and an emission wavelength of 525 nm. ( c – e ) SOD, CAT activities, and MDA content: SOD, CAT activities, and MDA content were measured using respective assay kits. Data are presented as mean ± SD values of three independent experiments. * p
Figure Legend Snippet: SAL reduces H 2 O 2 -induced ROS production and MDA formation and increases the activities of antioxidant enzymes in HUVECs. ( a ) HUVECs were treated with various concentrations of H 2 O 2 (0–500 μM) for 4 h, and cell viability was determined by MTT assay. ( b – e ) HUVECs were incubated without any intervention (control), SAL (0.1, 1, 10 μM) for 24 h, or 300 μM H 2 O 2 for 4 h, or 0.1, 1, 10 μM SAL for 24 h followed by 300 μM H 2 O 2 for an additional 4 h. ( b ) Intercellular ROS levels: Intercellular ROS levels were determined by DCFH-DA fluorescence. After various treatment, the cells were stained with 10 μM DCFH-DA in serum free medium for 30 min in the dark. Fluorescence intensity was recorded using a Synergy TM 4 Multi-Mode Microplate Reader at an excitation wavelength of 488 nm and an emission wavelength of 525 nm. ( c – e ) SOD, CAT activities, and MDA content: SOD, CAT activities, and MDA content were measured using respective assay kits. Data are presented as mean ± SD values of three independent experiments. * p

Techniques Used: Multiple Displacement Amplification, MTT Assay, Incubation, Fluorescence, Staining

Effects of SAL on H 2 O 2 -induced cytotoxicity and oxidative stress in Nrf2 knockdown HUVECs. ( a ) Cell viability analysis. HUVECs were preincubated with or without Nrf2 siRNA, then treated with 1 μM SAL for 24 h before being treated with 300 μM H 2 O 2 for another 24 h, or 300 μM H 2 O 2 for 24 h. Cell viability was determined using MTT assay. ( b – e ) HUVECs were preincubated with or without Nrf2 siRNA, then treated with 300 μM H 2 O 2 for 4 h, or 1 μM SAL for 24 h followed by 300 μM H 2 O 2 for an additional 4 h. ( b ) Intercellular ROS levels: Intercellular ROS levels were determined by DCF fluorescence. After various treatment, the cells were stained with 10 μM DCFH-DA in serum free medium for 30 min in the dark. Fluorescence intensity was recorded using a Synergy TM 4 Multi-Mode Microplate Reader. ( c – e ) SOD, CAT activities, and MDA content: SOD, CAT activities, and MDA content were measured using respective assay kits. Data are presented as mean ± SD values of three independent experiments. * p
Figure Legend Snippet: Effects of SAL on H 2 O 2 -induced cytotoxicity and oxidative stress in Nrf2 knockdown HUVECs. ( a ) Cell viability analysis. HUVECs were preincubated with or without Nrf2 siRNA, then treated with 1 μM SAL for 24 h before being treated with 300 μM H 2 O 2 for another 24 h, or 300 μM H 2 O 2 for 24 h. Cell viability was determined using MTT assay. ( b – e ) HUVECs were preincubated with or without Nrf2 siRNA, then treated with 300 μM H 2 O 2 for 4 h, or 1 μM SAL for 24 h followed by 300 μM H 2 O 2 for an additional 4 h. ( b ) Intercellular ROS levels: Intercellular ROS levels were determined by DCF fluorescence. After various treatment, the cells were stained with 10 μM DCFH-DA in serum free medium for 30 min in the dark. Fluorescence intensity was recorded using a Synergy TM 4 Multi-Mode Microplate Reader. ( c – e ) SOD, CAT activities, and MDA content: SOD, CAT activities, and MDA content were measured using respective assay kits. Data are presented as mean ± SD values of three independent experiments. * p

Techniques Used: MTT Assay, Fluorescence, Staining, Multiple Displacement Amplification

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