Structured Review

Millipore hla dr2
( A ) SDS-PAGE analysis (12%) of detergent solubilized <t>HLA-DR2</t> (DRB5*0101/DRB1*1501; left) or HLA-DR4 (DRB1*0401; right) where 5–8×10 9 of the Epstein Barr virus (EBV)-transformed B cell line HTC-LAN and BSM were lysed, respectively, and their affinity was purified using the immobilized monoclonal antibody L243. After concentration on a 30-kDa membrane, 670–700 g of the human leukocyte antigen (HLA)-II were obtained and stored in phosphate buffered saline at a concentration of 0.5 mg/mL. The purity of HLA-DR2 and HLA-DR4 were about 85%. ( B ) Competition binding of the MBP 82–98 peptide analogues in 10-fold, 20-fold, and 50-fold excess of AMCA-labeled MBP 83–99 , which is specific to the HLA-DR2b chain, was used as the peptide competitor, and ( C ) AMCA-labeled HA 306–318 , which is specific to the HLA-DR4, was used as the peptide competitor. All of the experiments were in triplicates, where ** p
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Images

1) Product Images from "Design of Linear and Cyclic Mutant Analogues of Dirucotide Peptide (MBP82–98) against Multiple Sclerosis: Conformational and Binding Studies to MHC Class II"

Article Title: Design of Linear and Cyclic Mutant Analogues of Dirucotide Peptide (MBP82–98) against Multiple Sclerosis: Conformational and Binding Studies to MHC Class II

Journal: Brain Sciences

doi: 10.3390/brainsci8120213

( A ) SDS-PAGE analysis (12%) of detergent solubilized HLA-DR2 (DRB5*0101/DRB1*1501; left) or HLA-DR4 (DRB1*0401; right) where 5–8×10 9 of the Epstein Barr virus (EBV)-transformed B cell line HTC-LAN and BSM were lysed, respectively, and their affinity was purified using the immobilized monoclonal antibody L243. After concentration on a 30-kDa membrane, 670–700 g of the human leukocyte antigen (HLA)-II were obtained and stored in phosphate buffered saline at a concentration of 0.5 mg/mL. The purity of HLA-DR2 and HLA-DR4 were about 85%. ( B ) Competition binding of the MBP 82–98 peptide analogues in 10-fold, 20-fold, and 50-fold excess of AMCA-labeled MBP 83–99 , which is specific to the HLA-DR2b chain, was used as the peptide competitor, and ( C ) AMCA-labeled HA 306–318 , which is specific to the HLA-DR4, was used as the peptide competitor. All of the experiments were in triplicates, where ** p
Figure Legend Snippet: ( A ) SDS-PAGE analysis (12%) of detergent solubilized HLA-DR2 (DRB5*0101/DRB1*1501; left) or HLA-DR4 (DRB1*0401; right) where 5–8×10 9 of the Epstein Barr virus (EBV)-transformed B cell line HTC-LAN and BSM were lysed, respectively, and their affinity was purified using the immobilized monoclonal antibody L243. After concentration on a 30-kDa membrane, 670–700 g of the human leukocyte antigen (HLA)-II were obtained and stored in phosphate buffered saline at a concentration of 0.5 mg/mL. The purity of HLA-DR2 and HLA-DR4 were about 85%. ( B ) Competition binding of the MBP 82–98 peptide analogues in 10-fold, 20-fold, and 50-fold excess of AMCA-labeled MBP 83–99 , which is specific to the HLA-DR2b chain, was used as the peptide competitor, and ( C ) AMCA-labeled HA 306–318 , which is specific to the HLA-DR4, was used as the peptide competitor. All of the experiments were in triplicates, where ** p

Techniques Used: SDS Page, Transformation Assay, Purification, Concentration Assay, Binding Assay, Labeling

2) Product Images from "Splice Variants of Perlucin from Haliotis laevigata Modulate the Crystallisation of CaCO3"

Article Title: Splice Variants of Perlucin from Haliotis laevigata Modulate the Crystallisation of CaCO3

Journal: PLoS ONE

doi: 10.1371/journal.pone.0097126

Amino acid sequences of Perlucin from H. laevigata . Perlucin splice variants (Perlucin-R0, Perlucin-R5, Perlucin-R8) are indicated as grey bars above the sequence. The following characteristics of the proteins are marked: Amino acid exchanges in Perlucin-R0 (M89I, V129D, R149L), signal peptide, C-type lectin domain [60] , repeat units (light blue bars). Peptides identified from 2D electrophoresis spots ( Figure 3 ) by MALDI-ToF MS (black bars) or ESI-MS (red bar). *Predicted glycosylation and phosphorylation (NetPhos [50] ) sites.
Figure Legend Snippet: Amino acid sequences of Perlucin from H. laevigata . Perlucin splice variants (Perlucin-R0, Perlucin-R5, Perlucin-R8) are indicated as grey bars above the sequence. The following characteristics of the proteins are marked: Amino acid exchanges in Perlucin-R0 (M89I, V129D, R149L), signal peptide, C-type lectin domain [60] , repeat units (light blue bars). Peptides identified from 2D electrophoresis spots ( Figure 3 ) by MALDI-ToF MS (black bars) or ESI-MS (red bar). *Predicted glycosylation and phosphorylation (NetPhos [50] ) sites.

Techniques Used: Sequencing, Two-Dimensional Gel Electrophoresis, Mass Spectrometry

Electrophoretic analysis of recombinant and native Perlucin preparations. A) Western blot of cell culture supernatants derived from COS-7 cells ectopically over expressing the indicated Strep-tagged recombinant Perlucins, which were detected using a polyclonal anti-Strep-tag antibody as described in the Methods section. B) SDS-PAGE of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie brilliant blue showed one distinct band at approx. 25 kDa, one at 20 kDa, and one at approx. 15 kDa. C) 2D electrophoresis of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie Brilliant Blue. The indicated spots were cut out and analysed by MALDI-ToF MS as described in the Methods section. Spot 7 was used as control.
Figure Legend Snippet: Electrophoretic analysis of recombinant and native Perlucin preparations. A) Western blot of cell culture supernatants derived from COS-7 cells ectopically over expressing the indicated Strep-tagged recombinant Perlucins, which were detected using a polyclonal anti-Strep-tag antibody as described in the Methods section. B) SDS-PAGE of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie brilliant blue showed one distinct band at approx. 25 kDa, one at 20 kDa, and one at approx. 15 kDa. C) 2D electrophoresis of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie Brilliant Blue. The indicated spots were cut out and analysed by MALDI-ToF MS as described in the Methods section. Spot 7 was used as control.

Techniques Used: Recombinant, Western Blot, Cell Culture, Derivative Assay, Expressing, Strep-tag, SDS Page, Purification, Staining, Two-Dimensional Gel Electrophoresis, Mass Spectrometry

3) Product Images from "Effect of Uncaria tomentosa extract on purinergic enzyme activities in lymphocytes of rats submitted to experimental adjuvant arthritis model"

Article Title: Effect of Uncaria tomentosa extract on purinergic enzyme activities in lymphocytes of rats submitted to experimental adjuvant arthritis model

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/s12906-015-0694-4

ATP ( a ) and ADP ( b ) hydrolysis in lymphocytes of Complete Freund’s Adjuvant (CFA)- induced arthritis rats and treated for 45 days with Uncaria tomentosa extract in the dose of 150 mg/kg, 2 times/day. Enzyme specific activities are reported as nmol of Pi released/min/mg of protein. Groups: C (control), E (extract), A (arthritis) and A + E (arthritis + extract). Bars represent mean ± S.E.M. ( a,b ) Indicates a significant P
Figure Legend Snippet: ATP ( a ) and ADP ( b ) hydrolysis in lymphocytes of Complete Freund’s Adjuvant (CFA)- induced arthritis rats and treated for 45 days with Uncaria tomentosa extract in the dose of 150 mg/kg, 2 times/day. Enzyme specific activities are reported as nmol of Pi released/min/mg of protein. Groups: C (control), E (extract), A (arthritis) and A + E (arthritis + extract). Bars represent mean ± S.E.M. ( a,b ) Indicates a significant P

Techniques Used:

4) Product Images from "Method for simultaneous analysis of eight analogues of vitamin D using liquid chromatography tandem mass spectrometry"

Article Title: Method for simultaneous analysis of eight analogues of vitamin D using liquid chromatography tandem mass spectrometry

Journal: Chemistry Central Journal

doi: 10.1186/1752-153X-6-112

Typical chromatogram showing vitamin D analogues [1 ST period analytes: Stanozolol-d3-RT = 2.96, 1α25(OH) 2 D2-RT = 5.49, 1α25(OH) 2 D3-RT = 6.74; 2 nd period analytes: 7αC4-RT = 9.51, 25OHD3-RT = 10.86, 3-epi-25OHD3-RT = 11.21, 25OHD2-RT = 11.46 and 3-epi-25OHD2-RT = 11.71; 3 RD period analytes: vitamin D3-RT = 14.87 and vitamin D2-RT = 15.38].
Figure Legend Snippet: Typical chromatogram showing vitamin D analogues [1 ST period analytes: Stanozolol-d3-RT = 2.96, 1α25(OH) 2 D2-RT = 5.49, 1α25(OH) 2 D3-RT = 6.74; 2 nd period analytes: 7αC4-RT = 9.51, 25OHD3-RT = 10.86, 3-epi-25OHD3-RT = 11.21, 25OHD2-RT = 11.46 and 3-epi-25OHD2-RT = 11.71; 3 RD period analytes: vitamin D3-RT = 14.87 and vitamin D2-RT = 15.38].

Techniques Used:

5) Product Images from "Square Wave Voltammetry of TNT at Gold Electrodes Modified with Self-Assembled Monolayers Containing Aromatic Structures"

Article Title: Square Wave Voltammetry of TNT at Gold Electrodes Modified with Self-Assembled Monolayers Containing Aromatic Structures

Journal: PLoS ONE

doi: 10.1371/journal.pone.0115966

Experimental details. A. Chemical structures for biphenyl-4-thiol (Biphenyl), 4-(phenylethynyl)benzenethiol (OPE) and undecane-1-thiol (C11) used to from SAMs on gold electrodes. B. The electrochemical setup.
Figure Legend Snippet: Experimental details. A. Chemical structures for biphenyl-4-thiol (Biphenyl), 4-(phenylethynyl)benzenethiol (OPE) and undecane-1-thiol (C11) used to from SAMs on gold electrodes. B. The electrochemical setup.

Techniques Used:

6) Product Images from "Haem-activated promiscuous targeting of artemisinin in Plasmodium falciparum"

Article Title: Haem-activated promiscuous targeting of artemisinin in Plasmodium falciparum

Journal: Nature Communications

doi: 10.1038/ncomms10111

In vitro binding and functional validation of artemisinin targets. ( a ) Artemisinin specifically interacts with OAT, PyrK, LDH, SpdSyn, SAMS and TCTP as the unlabelled artesunate (25 ×) can compete with the AP1 binding. Heat denaturation reduces the AP1 -labelling level of OAT, suggesting that the interaction of artemisinin with OAT is activity based. ( b ) Dose-dependent labelling of OAT with AP1 (4 h treatment). ( c ) Time-dependent labelling of OAT with AP1 . ( d ) The interaction of artemisinin with OAT may involve thiol and amine groups as IAA (blocking thiol, 30 mM) and NEM (blocking amine, 10 mM) pretreatment (20 min) can reduce binding. ( e , f ) Activated artesunate inhibits the activities of PyrK ( e ) and LDH ( f ) in vitro . Δ, heat denaturation; IAA, iodoacetamide; NEM, N -ethylmaleimide; Conc., concentration. Error bars represent s.d. in three independent replicates in e and f . Full-gel images for panels a – d are shown in Supplementary Fig. 13 .
Figure Legend Snippet: In vitro binding and functional validation of artemisinin targets. ( a ) Artemisinin specifically interacts with OAT, PyrK, LDH, SpdSyn, SAMS and TCTP as the unlabelled artesunate (25 ×) can compete with the AP1 binding. Heat denaturation reduces the AP1 -labelling level of OAT, suggesting that the interaction of artemisinin with OAT is activity based. ( b ) Dose-dependent labelling of OAT with AP1 (4 h treatment). ( c ) Time-dependent labelling of OAT with AP1 . ( d ) The interaction of artemisinin with OAT may involve thiol and amine groups as IAA (blocking thiol, 30 mM) and NEM (blocking amine, 10 mM) pretreatment (20 min) can reduce binding. ( e , f ) Activated artesunate inhibits the activities of PyrK ( e ) and LDH ( f ) in vitro . Δ, heat denaturation; IAA, iodoacetamide; NEM, N -ethylmaleimide; Conc., concentration. Error bars represent s.d. in three independent replicates in e and f . Full-gel images for panels a – d are shown in Supplementary Fig. 13 .

Techniques Used: In Vitro, Binding Assay, Functional Assay, Activity Assay, Blocking Assay, Concentration Assay

7) Product Images from "Interference of Quorum Sensing by Delftia sp. VM4 Depends on the Activity of a Novel N-Acylhomoserine Lactone-Acylase"

Article Title: Interference of Quorum Sensing by Delftia sp. VM4 Depends on the Activity of a Novel N-Acylhomoserine Lactone-Acylase

Journal: PLoS ONE

doi: 10.1371/journal.pone.0138034

Characterization of AHL-acylase of Delftia sp. VM4. (A) Silver stained 12% SDS-PAGE during purification of AHL acylase from Delftia sp. VM4. Lane: M, PMWH- protein marker; 1, ammonium sulphate precipitates; 2, eluate from DEAE-sepharose column; 3, eluate from Sephadex column. (B) Microtitre plate assay for the AHL degrading activity with 0.05 mM HHL of the renatured gel slices (bands i-vi) from the SDS-PAGE was assayed using C . violaceum CV026 biosensor based bioassay. HHL (0.05 mM) was loaded as the control (well-c). HPLC profile of (C) HHL without purified AHL acylase, (D) HHL degradation by purified AHL acylase, (E) EI-MS analysis of degraded product (HSL) from HHL.
Figure Legend Snippet: Characterization of AHL-acylase of Delftia sp. VM4. (A) Silver stained 12% SDS-PAGE during purification of AHL acylase from Delftia sp. VM4. Lane: M, PMWH- protein marker; 1, ammonium sulphate precipitates; 2, eluate from DEAE-sepharose column; 3, eluate from Sephadex column. (B) Microtitre plate assay for the AHL degrading activity with 0.05 mM HHL of the renatured gel slices (bands i-vi) from the SDS-PAGE was assayed using C . violaceum CV026 biosensor based bioassay. HHL (0.05 mM) was loaded as the control (well-c). HPLC profile of (C) HHL without purified AHL acylase, (D) HHL degradation by purified AHL acylase, (E) EI-MS analysis of degraded product (HSL) from HHL.

Techniques Used: Staining, SDS Page, Purification, Marker, Activity Assay, High Performance Liquid Chromatography, Mass Spectrometry

8) Product Images from "Localization of tamoxifen in human breast cancer tumors by MALDI mass spectrometry imaging"

Article Title: Localization of tamoxifen in human breast cancer tumors by MALDI mass spectrometry imaging

Journal: Clinical and Translational Medicine

doi: 10.1186/s40169-016-0090-9

Ionization characteristics of tamoxifen (0.1 mg/mL in water) as measured with 3.5 mg/mL CHCA on a stainless steel MALDI target plate. a A full mass spectrum of tamoxifen obtained at 60,000 resolution using the Orbitrap mass analyzer and b a tandem mass spectrum of tamoxifen isolating the m/z 372.23 and CID fragmented in the linear ion trap mass analyzer
Figure Legend Snippet: Ionization characteristics of tamoxifen (0.1 mg/mL in water) as measured with 3.5 mg/mL CHCA on a stainless steel MALDI target plate. a A full mass spectrum of tamoxifen obtained at 60,000 resolution using the Orbitrap mass analyzer and b a tandem mass spectrum of tamoxifen isolating the m/z 372.23 and CID fragmented in the linear ion trap mass analyzer

Techniques Used:

9) Product Images from "Macrophage receptors of polysaccharide isolated from a marine filamentous fungus Phoma herbarum YS4108"

Article Title: Macrophage receptors of polysaccharide isolated from a marine filamentous fungus Phoma herbarum YS4108

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2009.93

Polysaccharide labeling with fluoresceinamine. CDAP-activated YCP reacted with fluoresceinamine for 18 h at room temperature. The mixtures were fractionated on a Sephacryl S-400 column, concentration (μg/mL) of fluoresceinamine (▴) and YCP (□) in each fraction were determined. n =3.
Figure Legend Snippet: Polysaccharide labeling with fluoresceinamine. CDAP-activated YCP reacted with fluoresceinamine for 18 h at room temperature. The mixtures were fractionated on a Sephacryl S-400 column, concentration (μg/mL) of fluoresceinamine (▴) and YCP (□) in each fraction were determined. n =3.

Techniques Used: Labeling, Concentration Assay

10) Product Images from "Simultaneous Determination of Salidroside and Its Aglycone Metabolite p-Tyrosol in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry"

Article Title: Simultaneous Determination of Salidroside and Its Aglycone Metabolite p-Tyrosol in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry

Journal: Molecules

doi: 10.3390/molecules17044733

MRM chromatograms of salidroside, p -tyrosol and IS in ( a ) a blank rat plasma sample; ( b ) a blank rat plasma spiked with salidroside (500 ng·mL −1 ), p -tyrosol (100 ng·mL −1 ) and IS (200 ng·mL −1 ); ( c ) a rat plasma sample collected 5 min after i.v. administration of salidroside (50 mg/kg) with IS (200 ng·mL −1 ); ( d ) a rat plasma sample collected 5 min after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ); ( e ) a rat plasma sample collected 30 min after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ); ( f ) a rat plasma sample collected 2 h after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ); ( g ) a rat plasma sample collected 4 h after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ).
Figure Legend Snippet: MRM chromatograms of salidroside, p -tyrosol and IS in ( a ) a blank rat plasma sample; ( b ) a blank rat plasma spiked with salidroside (500 ng·mL −1 ), p -tyrosol (100 ng·mL −1 ) and IS (200 ng·mL −1 ); ( c ) a rat plasma sample collected 5 min after i.v. administration of salidroside (50 mg/kg) with IS (200 ng·mL −1 ); ( d ) a rat plasma sample collected 5 min after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ); ( e ) a rat plasma sample collected 30 min after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ); ( f ) a rat plasma sample collected 2 h after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ); ( g ) a rat plasma sample collected 4 h after i.g. administration of salidroside (100 mg/kg) with IS (200 ng·mL −1 ).

Techniques Used:

Product ion mass spectra of [M−H] − . ( a ) Salidroside ([M−H] − , m/z 299.0); ( b ) p- Tyrosol ([M−H] − , m/z 137.0); ( c ) IS (paracetamol) ([M−H] − , m/z 150.1).
Figure Legend Snippet: Product ion mass spectra of [M−H] − . ( a ) Salidroside ([M−H] − , m/z 299.0); ( b ) p- Tyrosol ([M−H] − , m/z 137.0); ( c ) IS (paracetamol) ([M−H] − , m/z 150.1).

Techniques Used:

Mean concentration-time profiles in rat plasma (n = 6) obtained after i.v. administration of salidroside (i.v. 50 mg/kg) and i.g. administration of salidroside (i.g. 100 mg/kg). ( a : salidroside; b : p- tyrosol).
Figure Legend Snippet: Mean concentration-time profiles in rat plasma (n = 6) obtained after i.v. administration of salidroside (i.v. 50 mg/kg) and i.g. administration of salidroside (i.g. 100 mg/kg). ( a : salidroside; b : p- tyrosol).

Techniques Used: Concentration Assay

Representative chromatograms of: ( a ) a blank rat plasma sample; ( b ) a blank rat plasma spiked with salidroside and p- tyrosol (20 μg/mL); ( c ) a rat plasma sample collected 5 min after i.v. administration of salidroside (50 mg/kg); ( d ) a rat plasma sample collected 30 min after i.g. administration of salidroside (100 mg/kg). (1: salidroside, 2: p -tyrosol).
Figure Legend Snippet: Representative chromatograms of: ( a ) a blank rat plasma sample; ( b ) a blank rat plasma spiked with salidroside and p- tyrosol (20 μg/mL); ( c ) a rat plasma sample collected 5 min after i.v. administration of salidroside (50 mg/kg); ( d ) a rat plasma sample collected 30 min after i.g. administration of salidroside (100 mg/kg). (1: salidroside, 2: p -tyrosol).

Techniques Used:

Chemical structures of ( a ) salidroside; ( b ) p- tyrosol; and ( c ) paracetamol (IS).
Figure Legend Snippet: Chemical structures of ( a ) salidroside; ( b ) p- tyrosol; and ( c ) paracetamol (IS).

Techniques Used:

11) Product Images from "Plant Defensive β-Glucosidases Resist Digestion and Sustain Activity in the Gut of a Lepidopteran Herbivore"

Article Title: Plant Defensive β-Glucosidases Resist Digestion and Sustain Activity in the Gut of a Lepidopteran Herbivore

Journal: Frontiers in Plant Science

doi: 10.3389/fpls.2018.01389

Maize β-glucosidases were recovered in active form after digestion by S. littoralis larvae. Enzymatic activity levels were determined in non-ingested material and after digestion of diet cubes spiked with semi-purified maize β-glucosidase (A) , or after feeding on maize leaf tissue (B) . Activities were measured either using the non-specific β-glucosidase substrate pNPG (A) or the endogenous plant substrate DIMBOA-Glc (B) . Shown are the means ± standard errors [ N = 3 in both analyses; P = 0.2 (A) and P = 0.3 (B) ].
Figure Legend Snippet: Maize β-glucosidases were recovered in active form after digestion by S. littoralis larvae. Enzymatic activity levels were determined in non-ingested material and after digestion of diet cubes spiked with semi-purified maize β-glucosidase (A) , or after feeding on maize leaf tissue (B) . Activities were measured either using the non-specific β-glucosidase substrate pNPG (A) or the endogenous plant substrate DIMBOA-Glc (B) . Shown are the means ± standard errors [ N = 3 in both analyses; P = 0.2 (A) and P = 0.3 (B) ].

Techniques Used: Activity Assay, Purification, Gas Chromatography

Schematic representation of some reactions catalyzed by plant β-glucosidases. (A) Activation of glucosinolates by myrosinase. (B) Step-wise hydrolysis of the cyanogenic diglucoside amygdalin. (C) Hydrolytic activation of the benzoxazinoid glucoside DIMBOA-Glc and formation of the open-ring aglycone form in solution.
Figure Legend Snippet: Schematic representation of some reactions catalyzed by plant β-glucosidases. (A) Activation of glucosinolates by myrosinase. (B) Step-wise hydrolysis of the cyanogenic diglucoside amygdalin. (C) Hydrolytic activation of the benzoxazinoid glucoside DIMBOA-Glc and formation of the open-ring aglycone form in solution.

Techniques Used: Activation Assay, Gas Chromatography

12) Product Images from "The Metabolism of Salidroside to Its Aglycone p-Tyrosol in Rats following the Administration of Salidroside"

Article Title: The Metabolism of Salidroside to Its Aglycone p-Tyrosol in Rats following the Administration of Salidroside

Journal: PLoS ONE

doi: 10.1371/journal.pone.0103648

Mean concentration-time profiles of (A) salidroside and (B) p- tyrosol in rat tissues (n = 6) obtained after i.v. administration of salidroside (i.v. 50 mg/kg).
Figure Legend Snippet: Mean concentration-time profiles of (A) salidroside and (B) p- tyrosol in rat tissues (n = 6) obtained after i.v. administration of salidroside (i.v. 50 mg/kg).

Techniques Used: Concentration Assay

Mean concentration-time profiles of (A) salidroside and (B) p- tyrosol in rat tissues (n = 6) obtained after i.g. administration of salidroside (i.g. 100 mg/kg).
Figure Legend Snippet: Mean concentration-time profiles of (A) salidroside and (B) p- tyrosol in rat tissues (n = 6) obtained after i.g. administration of salidroside (i.g. 100 mg/kg).

Techniques Used: Concentration Assay

Chemical structures of (A) salidroside, (B) p- tyrosol and (C) paracetamol (IS).
Figure Legend Snippet: Chemical structures of (A) salidroside, (B) p- tyrosol and (C) paracetamol (IS).

Techniques Used:

MRM chromatograms of salidroside, p -tyrosol and the IS in (A) a blank rat liver tissue homogenate sample, (B) a blank rat liver tissue sample spiked with salidroside (500 ng/mL), p -tyrosol (500 ng/mL) and the IS (200 ng/mL), (C) a rat liver tissue homogenate sample collected 0.17 h after i.v. administration of salidroside (50 mg/kg) with the IS (200 ng/mL), (D) a rat liver tissue homogenate sample collected 1 h after i.g. administration of salidroside (100 mg/kg) with the IS (200 ng/mL).
Figure Legend Snippet: MRM chromatograms of salidroside, p -tyrosol and the IS in (A) a blank rat liver tissue homogenate sample, (B) a blank rat liver tissue sample spiked with salidroside (500 ng/mL), p -tyrosol (500 ng/mL) and the IS (200 ng/mL), (C) a rat liver tissue homogenate sample collected 0.17 h after i.v. administration of salidroside (50 mg/kg) with the IS (200 ng/mL), (D) a rat liver tissue homogenate sample collected 1 h after i.g. administration of salidroside (100 mg/kg) with the IS (200 ng/mL).

Techniques Used:

13) Product Images from "A subcortical inhibitory signal for behavioral arrest in the thalamus"

Article Title: A subcortical inhibitory signal for behavioral arrest in the thalamus

Journal: Nature neuroscience

doi: 10.1038/nn.3951

Activation of glycinergic afferents interrupts ongoing cortical activity. a) Representative standardized frontal cortical LFP traces before (1) and during (2) the optogenetic activation of GlyT2 fibers in the IL. b) Wavelet spectrum of the cortical LFP showing the 33 s long activation period together with pre- and post-illumination period. Grey bars indicate the position of LFP samples in (a). Warm colors indicate higher power. c) Power spectra of the cortical LFPs in the 30 s preceding the stimulation (orange) and during photoactivation (blue) of the GlyT2 fibers in IL. d) Statistical comparison of the power spectra of the stimulated and control periods in one representative animal (n=25 stimulations). Gray bar indicates the frequency range which displayed statistically significant difference (2.14 Hz - 5.8 Hz, Mann-Whitney U test). In this range the highest p value was 0.00226 at 5.8 Hz (W=457). All other p values were lower. Error bars represent the s.e.m. au arbitrary unit.
Figure Legend Snippet: Activation of glycinergic afferents interrupts ongoing cortical activity. a) Representative standardized frontal cortical LFP traces before (1) and during (2) the optogenetic activation of GlyT2 fibers in the IL. b) Wavelet spectrum of the cortical LFP showing the 33 s long activation period together with pre- and post-illumination period. Grey bars indicate the position of LFP samples in (a). Warm colors indicate higher power. c) Power spectra of the cortical LFPs in the 30 s preceding the stimulation (orange) and during photoactivation (blue) of the GlyT2 fibers in IL. d) Statistical comparison of the power spectra of the stimulated and control periods in one representative animal (n=25 stimulations). Gray bar indicates the frequency range which displayed statistically significant difference (2.14 Hz - 5.8 Hz, Mann-Whitney U test). In this range the highest p value was 0.00226 at 5.8 Hz (W=457). All other p values were lower. Error bars represent the s.e.m. au arbitrary unit.

Techniques Used: Activation Assay, Activity Assay, MANN-WHITNEY

Glycinergic terminals in IL are multisynaptic, co-express GABA and display variable postsynaptic receptor composition. a) 3D reconstruction of a GlyT2::eGFP -positive terminal in IL from serial electron microscopic (EM) images, three of which are shown on the right. Green, synapses; magenta, puncta adherentia; dark blue, membrane of the terminal; light blue, glia; arrows, synapse; arrowheads puncta adherantia. b) Consecutive electron micrographs of a GlyT2:eGFP bouton (b) in the mouse IL immunostained for eGFP using preembedding silver staining (left), and for GABA using postembedding immunogold labeling (right). c) Comparison of random dendritic diameters (white bars) in the IL and the diameter of targets postsynaptic to GlyT2::eGFP terminals (black bars). Random dendrite diameters are also shown and as the ratio of summated perimeter of the dendrites in each bin (black line with diamonds), which better reflect the available membrane surfaces. d) Correlation between the synapse numbers of the GlyT2::eGFP boutons and the diameter of the postsynaptic IL dendrites. e) The average number of synapses with increasing distances from a given synapse in eGFP boutons in the IL. f) Electron micrograph of a GlyT2-immunopositive axon terminal in the human IL. green arrowheads, synapses g) White arrows point to colocalization of the γ2 subunit of GABA A receptors and of glycine receptors postsynaptic to GlyT2::eGFP terminals. The cityscape plot (h) represents the number of apposed GABAγ2R receptor and GlyR clusters per GlyT2::eGFP varicosity. Scales: a, b, f, 500 nm; g 1μm.
Figure Legend Snippet: Glycinergic terminals in IL are multisynaptic, co-express GABA and display variable postsynaptic receptor composition. a) 3D reconstruction of a GlyT2::eGFP -positive terminal in IL from serial electron microscopic (EM) images, three of which are shown on the right. Green, synapses; magenta, puncta adherentia; dark blue, membrane of the terminal; light blue, glia; arrows, synapse; arrowheads puncta adherantia. b) Consecutive electron micrographs of a GlyT2:eGFP bouton (b) in the mouse IL immunostained for eGFP using preembedding silver staining (left), and for GABA using postembedding immunogold labeling (right). c) Comparison of random dendritic diameters (white bars) in the IL and the diameter of targets postsynaptic to GlyT2::eGFP terminals (black bars). Random dendrite diameters are also shown and as the ratio of summated perimeter of the dendrites in each bin (black line with diamonds), which better reflect the available membrane surfaces. d) Correlation between the synapse numbers of the GlyT2::eGFP boutons and the diameter of the postsynaptic IL dendrites. e) The average number of synapses with increasing distances from a given synapse in eGFP boutons in the IL. f) Electron micrograph of a GlyT2-immunopositive axon terminal in the human IL. green arrowheads, synapses g) White arrows point to colocalization of the γ2 subunit of GABA A receptors and of glycine receptors postsynaptic to GlyT2::eGFP terminals. The cityscape plot (h) represents the number of apposed GABAγ2R receptor and GlyR clusters per GlyT2::eGFP varicosity. Scales: a, b, f, 500 nm; g 1μm.

Techniques Used: Silver Staining, Labeling

Glycinergic input evokes non-depressing inhibition and reduces IL cell firing. a) Scheme of the experiment. b) ChR2-eYFP containing fibers in the IL. c) Averaged sample trace of light-evoked IPSCs before (black trace) and after (red) application of gabazine and strychnine. d) Variable mixed GABA/glycinergic phenotype of light-evoked IPSCs. Three different examples are shown with only GABAergic (top), mixed GABA/glycinergic (center), and only glycinergic (bottom) transmission. e) Ratio of IL cells showing various proportions of glycinergic leIPSCs. f-g ) SR95531 application leads to a significant acceleration of the decay time course of the leIPSCs. See the averaged traces for a single recorded cell in (f), and the pooled results for all the experiments in (g). h) Light-evoked responses display little depression during stimulation trains at different frequencies. i) Activation of GlyT2 fibers interrupts firing of IL neurons recorded in the current clamp configuration.
Figure Legend Snippet: Glycinergic input evokes non-depressing inhibition and reduces IL cell firing. a) Scheme of the experiment. b) ChR2-eYFP containing fibers in the IL. c) Averaged sample trace of light-evoked IPSCs before (black trace) and after (red) application of gabazine and strychnine. d) Variable mixed GABA/glycinergic phenotype of light-evoked IPSCs. Three different examples are shown with only GABAergic (top), mixed GABA/glycinergic (center), and only glycinergic (bottom) transmission. e) Ratio of IL cells showing various proportions of glycinergic leIPSCs. f-g ) SR95531 application leads to a significant acceleration of the decay time course of the leIPSCs. See the averaged traces for a single recorded cell in (f), and the pooled results for all the experiments in (g). h) Light-evoked responses display little depression during stimulation trains at different frequencies. i) Activation of GlyT2 fibers interrupts firing of IL neurons recorded in the current clamp configuration.

Techniques Used: Inhibition, Transmission Assay, Activation Assay

Activation of glycinergic afferents interrupts ongoing behavior activity. a) Experimental design. b) Mice trajectory during the 1 st (red) and 2 nd (green) 5 s of optogenetic activation of GlyT2 fibers in the IL and during laser light shut off (black lines), in control (eYFP, left) and experimental (ChR2-eYFP, right) conditions. c) Average movement of control (dashed trace) and optogenetically activated (continuous trace) mice before, during (blue bar) and after stimulation. Error bars represent the s.e.m.
Figure Legend Snippet: Activation of glycinergic afferents interrupts ongoing behavior activity. a) Experimental design. b) Mice trajectory during the 1 st (red) and 2 nd (green) 5 s of optogenetic activation of GlyT2 fibers in the IL and during laser light shut off (black lines), in control (eYFP, left) and experimental (ChR2-eYFP, right) conditions. c) Average movement of control (dashed trace) and optogenetically activated (continuous trace) mice before, during (blue bar) and after stimulation. Error bars represent the s.e.m.

Techniques Used: Activation Assay, Activity Assay, Mouse Assay

Activity of GlyT2-positive neurons in the PRF in vivo is linked to cortical slow oscillation. a) Experimental design. b) Spiking activity of a GlyT2 cell in vivo under ketamine-xylazine anesthesia (bottom trace) together with the cortical LFP (top trace) and filtered cortical multiunit activity (MUA, middle trace). c) The recorded and neurobiotin filled cell display GlyT2::eGFP expression. d) Neurolucida reconstruction of the recorded cell . e-f) Phase distribution of the firing activity of five different GlyT2::eGFP -positive neurons relative to the cortical slow oscillation. One cycle is 360 o , 0 o peak of the UP state. Note the different phase preference of each cell. Scale: c, 20 µm; d, 100 µm.
Figure Legend Snippet: Activity of GlyT2-positive neurons in the PRF in vivo is linked to cortical slow oscillation. a) Experimental design. b) Spiking activity of a GlyT2 cell in vivo under ketamine-xylazine anesthesia (bottom trace) together with the cortical LFP (top trace) and filtered cortical multiunit activity (MUA, middle trace). c) The recorded and neurobiotin filled cell display GlyT2::eGFP expression. d) Neurolucida reconstruction of the recorded cell . e-f) Phase distribution of the firing activity of five different GlyT2::eGFP -positive neurons relative to the cortical slow oscillation. One cycle is 360 o , 0 o peak of the UP state. Note the different phase preference of each cell. Scale: c, 20 µm; d, 100 µm.

Techniques Used: Activity Assay, In Vivo, Expressing

Glycinergic afferents in the mouse and human IL. a) Injection site of the retrograde tracer fluorogold (FG) into the IL of a GlyT2::eGFP mouse. b-d) Retrogradely labeled GlyT2::eGFP -positive (arrows) and negative (arrowheads) neurons in the nucleus pontis oralis (PnO) at the coronal level indicated in the inset. e) Injection site of the anterograde tracer PHAL into the PnO and ( f-g) anterogradely labeled GlyT2::eGFP -positive fibers (arrows) in the IL at the position shown in the inset. Distribution of GlyT2 fibers in the mouse ( i,j) and human ( m,n) thalamus at two coronal levels. The figures represent cumulative data. Light microscopic images of GlyT2-positive fibers and innervation of calbindin-positive cells via multiple contacts in the mouse (k, l) and human ( o, p) IL. Scale bars: A,E, 1mm; all other 20 µm. CB, calbindin, For other abbreviations see Supplementary Fig 1,2 .
Figure Legend Snippet: Glycinergic afferents in the mouse and human IL. a) Injection site of the retrograde tracer fluorogold (FG) into the IL of a GlyT2::eGFP mouse. b-d) Retrogradely labeled GlyT2::eGFP -positive (arrows) and negative (arrowheads) neurons in the nucleus pontis oralis (PnO) at the coronal level indicated in the inset. e) Injection site of the anterograde tracer PHAL into the PnO and ( f-g) anterogradely labeled GlyT2::eGFP -positive fibers (arrows) in the IL at the position shown in the inset. Distribution of GlyT2 fibers in the mouse ( i,j) and human ( m,n) thalamus at two coronal levels. The figures represent cumulative data. Light microscopic images of GlyT2-positive fibers and innervation of calbindin-positive cells via multiple contacts in the mouse (k, l) and human ( o, p) IL. Scale bars: A,E, 1mm; all other 20 µm. CB, calbindin, For other abbreviations see Supplementary Fig 1,2 .

Techniques Used: Injection, Labeling

14) Product Images from "MASTL overexpression promotes chromosome instability and metastasis in breast cancer"

Article Title: MASTL overexpression promotes chromosome instability and metastasis in breast cancer

Journal: Oncogene

doi: 10.1038/s41388-018-0295-z

MASTL overexpression results in the loss of cell–cell junctions, causing migration defects. a Representative maximum projection images from confocal immunofluorescence of control and MASTL stained with H33342 (cyan) for E-Cadherin (green), β-catenin (yellow), and phalloidin (F-actin, pink). Scale bar 10 µm. b Wound-healing assays of control, and MASTL cell lines. Representative phase contrast, with inset of invading cell front (Control) and individual cells (MASTL) shown. Scale bar 50 µm. Kymographs (mCherry) generated by horizontal line through middle of wound area. Yellow dotted line indicates wound closure. c MRI Wound-Healing Tool for ImageJ was used to determine unclosed wound area as a function of time (mean ± SEM shown). Dotted line indicates 50% closure. Trend-line analysis performed using asymmetric sigmoidal analysis in Prism ( R 2 = 0.5888; Control and 0.7912; MASTL). d The MTrackJ plugin for ImageJ was used to track individual cells ( n = minimum 50 cell/condition) from b . e Data from d were analysed using the DiPer software tool. Shown are average speed and directionality ratios for MASTL and controls from three independent experiments (mean ± SEM, unpaired t -test, **** p
Figure Legend Snippet: MASTL overexpression results in the loss of cell–cell junctions, causing migration defects. a Representative maximum projection images from confocal immunofluorescence of control and MASTL stained with H33342 (cyan) for E-Cadherin (green), β-catenin (yellow), and phalloidin (F-actin, pink). Scale bar 10 µm. b Wound-healing assays of control, and MASTL cell lines. Representative phase contrast, with inset of invading cell front (Control) and individual cells (MASTL) shown. Scale bar 50 µm. Kymographs (mCherry) generated by horizontal line through middle of wound area. Yellow dotted line indicates wound closure. c MRI Wound-Healing Tool for ImageJ was used to determine unclosed wound area as a function of time (mean ± SEM shown). Dotted line indicates 50% closure. Trend-line analysis performed using asymmetric sigmoidal analysis in Prism ( R 2 = 0.5888; Control and 0.7912; MASTL). d The MTrackJ plugin for ImageJ was used to track individual cells ( n = minimum 50 cell/condition) from b . e Data from d were analysed using the DiPer software tool. Shown are average speed and directionality ratios for MASTL and controls from three independent experiments (mean ± SEM, unpaired t -test, **** p

Techniques Used: Over Expression, Migration, Immunofluorescence, Staining, Generated, Magnetic Resonance Imaging, Software

15) Product Images from "Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle"

Article Title: Removal of Abnormal Myofilament O-GlcNAcylation Restores Ca2+ Sensitivity in Diabetic Cardiac Muscle

Journal: Diabetes

doi: 10.2337/db14-1107

Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.
Figure Legend Snippet: Differential OGT and OGA subcellular localization and myofilament interactions in control and diabetic myocardium. Representative transmission electron microscopy images of control ( A ) and STZ diabetic rat myocardium ( B ). Ultrathin sections were examined with immunoelectron microscopy with primary antibodies (anti-OGT AL-28 and anti-OGA), and secondary antibodies gold-labeled (anti-rabbit, 12 nm; anti-chicken, 6 nm). Z-line, green arrowhead; OGT, purple circles; OGA, red circles; OGT and OGA in close vicinity, pink circles; OGT, purple arrowhead; and OGA, red arrowhead. Quantification of OGT ( C ) and OGA ( D ) number of particles/field in nine fields shows an increase for OGT (2 ± 0.6 vs. 8.6 ± 2.6, * P ≤ 0.028) and OGA (9.4 ± 2.9 vs 37.6 ± 12.6, * P ≤ 0.05) immunoelectron microscopy in STZ diabetic myocardium. Images were analyzed in ImageJ software (NIH). Representative coimmunoprecipitations (co-IP) of OGT ( E ) and OGA ( F ), followed by Western blots (WB) for α-sarcomeric actin, Tm, and MLC 1. A fraction of the inputs from controls (C1, C2), STZ (S1, S2), and agarose beads with no antibody (M) or isotype-specific normal antibody + agarose beads (1°) were included. G : Analysis of integrated signal density of myofilament immunoreactivity normalized to total IP OGT displayed a trend toward increased interactions with Tm and MLC 1. H : OGA co-IP shows that diabetic STZ rats display several fold increased associations with α-actin, α-Tm, and MLC 1. * P ≤ 0.05.

Techniques Used: Transmission Assay, Electron Microscopy, Immuno-Electron Microscopy, Labeling, Software, Co-Immunoprecipitation Assay, Western Blot

16) Product Images from "ELTA: Enzymatic Labeling of Terminal ADP-ribose"

Article Title: ELTA: Enzymatic Labeling of Terminal ADP-ribose

Journal: Molecular cell

doi: 10.1016/j.molcel.2018.12.022

ELTA labels free or protein-conjugated ADP-ribose monomers and polymers. (a) Schematics of ELTA. Free or protein-conjugated ADP-ribose can be labeled by incubating with OAS1 and dATP, where the 2’-OH terminus is indicated in red. Colored box indicates various dATP analogs that can also be used in the ELTA reactions, including radioactive ( 32 P), fluorescent (Cy3, Cy5), biotinylated or clickable analogs. (b) 15% urea-PAGE analyses of the addition of 32 P-dAMP onto ADP-ribose monomers and polymers using ELTA and visualized by autoradiograph. (c) MALDI-TOF analyses of the reaction of ADP-ribose with dATP, and with or without OAS1. (d-e) Analyses of the ELTA labeling reaction of (d) MARylated PARP10 catalytic domain (mod-PARP10 cd ) and (e) PARylated ha PARP (mod- ha PARP) using 32 P-dATP. Shown are a coomassie gel (left), an autoradiograph (middle), and a western blot probed with pan-ADP-ribose reagent (right). As negative controls, modified proteins were treated with the phosphodiesterase hs NudT16 to remove the 2’-OH termini of the ADP-ribose groups prior to ELTA labeling. For panel d, * indicates PARP10; OAS1 was ADP-ribosylated by PARP10 with the remnant of NAD + , and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1. For panel e, * indicates ha PARP and § indicates ha PARP fragments that were also ADP-ribosylated and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1.
Figure Legend Snippet: ELTA labels free or protein-conjugated ADP-ribose monomers and polymers. (a) Schematics of ELTA. Free or protein-conjugated ADP-ribose can be labeled by incubating with OAS1 and dATP, where the 2’-OH terminus is indicated in red. Colored box indicates various dATP analogs that can also be used in the ELTA reactions, including radioactive ( 32 P), fluorescent (Cy3, Cy5), biotinylated or clickable analogs. (b) 15% urea-PAGE analyses of the addition of 32 P-dAMP onto ADP-ribose monomers and polymers using ELTA and visualized by autoradiograph. (c) MALDI-TOF analyses of the reaction of ADP-ribose with dATP, and with or without OAS1. (d-e) Analyses of the ELTA labeling reaction of (d) MARylated PARP10 catalytic domain (mod-PARP10 cd ) and (e) PARylated ha PARP (mod- ha PARP) using 32 P-dATP. Shown are a coomassie gel (left), an autoradiograph (middle), and a western blot probed with pan-ADP-ribose reagent (right). As negative controls, modified proteins were treated with the phosphodiesterase hs NudT16 to remove the 2’-OH termini of the ADP-ribose groups prior to ELTA labeling. For panel d, * indicates PARP10; OAS1 was ADP-ribosylated by PARP10 with the remnant of NAD + , and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1. For panel e, * indicates ha PARP and § indicates ha PARP fragments that were also ADP-ribosylated and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1.

Techniques Used: Labeling, Polyacrylamide Gel Electrophoresis, Autoradiography, Western Blot, Modification

17) Product Images from "ELTA: Enzymatic Labeling of Terminal ADP-ribose"

Article Title: ELTA: Enzymatic Labeling of Terminal ADP-ribose

Journal: Molecular cell

doi: 10.1016/j.molcel.2018.12.022

ELTA labels free or protein-conjugated ADP-ribose monomers and polymers. (a) Schematics of ELTA. Free or protein-conjugated ADP-ribose can be labeled by incubating with OAS1 and dATP, where the 2’-OH terminus is indicated in red. Colored box indicates various dATP analogs that can also be used in the ELTA reactions, including radioactive ( 32 P), fluorescent (Cy3, Cy5), biotinylated or clickable analogs. (b) 15% urea-PAGE analyses of the addition of 32 P-dAMP onto ADP-ribose monomers and polymers using ELTA and visualized by autoradiograph. (c) MALDI-TOF analyses of the reaction of ADP-ribose with dATP, and with or without OAS1. (d-e) Analyses of the ELTA labeling reaction of (d) MARylated PARP10 catalytic domain (mod-PARP10 cd ) and (e) PARylated ha PARP (mod- ha PARP) using 32 P-dATP. Shown are a coomassie gel (left), an autoradiograph (middle), and a western blot probed with pan-ADP-ribose reagent (right). As negative controls, modified proteins were treated with the phosphodiesterase hs NudT16 to remove the 2’-OH termini of the ADP-ribose groups prior to ELTA labeling. For panel d, * indicates PARP10; OAS1 was ADP-ribosylated by PARP10 with the remnant of NAD + , and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1. For panel e, * indicates ha PARP and § indicates ha PARP fragments that were also ADP-ribosylated and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1.
Figure Legend Snippet: ELTA labels free or protein-conjugated ADP-ribose monomers and polymers. (a) Schematics of ELTA. Free or protein-conjugated ADP-ribose can be labeled by incubating with OAS1 and dATP, where the 2’-OH terminus is indicated in red. Colored box indicates various dATP analogs that can also be used in the ELTA reactions, including radioactive ( 32 P), fluorescent (Cy3, Cy5), biotinylated or clickable analogs. (b) 15% urea-PAGE analyses of the addition of 32 P-dAMP onto ADP-ribose monomers and polymers using ELTA and visualized by autoradiograph. (c) MALDI-TOF analyses of the reaction of ADP-ribose with dATP, and with or without OAS1. (d-e) Analyses of the ELTA labeling reaction of (d) MARylated PARP10 catalytic domain (mod-PARP10 cd ) and (e) PARylated ha PARP (mod- ha PARP) using 32 P-dATP. Shown are a coomassie gel (left), an autoradiograph (middle), and a western blot probed with pan-ADP-ribose reagent (right). As negative controls, modified proteins were treated with the phosphodiesterase hs NudT16 to remove the 2’-OH termini of the ADP-ribose groups prior to ELTA labeling. For panel d, * indicates PARP10; OAS1 was ADP-ribosylated by PARP10 with the remnant of NAD + , and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1. For panel e, * indicates ha PARP and § indicates ha PARP fragments that were also ADP-ribosylated and, therefore, detected by pan-ADP-ribose reagent and labeled by OAS1.

Techniques Used: Labeling, Polyacrylamide Gel Electrophoresis, Autoradiography, Western Blot, Modification

Detection of ADP-ribose length from individual proteins and cells using ELTA. (a) 15% urea-PAGE analyses of the ELTA labeling reaction of ADP-ribose monomer (lane 1) and PAR of mixed length (lane 2), as well as ADP-ribose monomers and polymers isolated from PARP1 automodification reactions with 1 mM NAD + for 0 (lane 3), 10 (lane 4), or 30 min (lane 5) that were labeled by OAS1 and 32 P-dATP. As a comparison, the ADP-ribose isolated from PARP1 automodification reaction in the same time frame with 1 mM NAD + with a trace of 32 P-NAD + were loaded in lanes 6–8. We note that 50-fold less of the reaction were loaded in lanes 3–5 compared with lanes 6–8. (b) 15% urea-PAGE analyses of ELTA labeling reaction of ADP-ribose monomers and polymers isolated from automodification of PARP1 along with either BSA and HPF1. The first lane contained ELTA-labeling of an equal mole of 5-, 10- and 20-mer PAR. The reaction in the PARP1+BSA lane was diluted 15 times in water prior to ELTA labeling. (c) 15% urea-PAGE analyses of the ELTA labeling reaction of ADP-ribose isolated from in vitro modified ha PARP (lane 1), from untreated HaCaT cells (lane 2), from HaCaT cells treated with 1 mM H 2 O 2 for 10 min (lane 3), from HaCaT cells treated with 1 mM H 2 O 2 for 10 min, but pre-treated the cells with 20 μM PARP inhibitor Olaparib for 2 h (lane 4), or pre-treated with 1 μM PARG inhibitor PDD00017273 for 2 h (lane 5). Corresponding lysates of cells from lanes 2–5 were probed with β-actin.
Figure Legend Snippet: Detection of ADP-ribose length from individual proteins and cells using ELTA. (a) 15% urea-PAGE analyses of the ELTA labeling reaction of ADP-ribose monomer (lane 1) and PAR of mixed length (lane 2), as well as ADP-ribose monomers and polymers isolated from PARP1 automodification reactions with 1 mM NAD + for 0 (lane 3), 10 (lane 4), or 30 min (lane 5) that were labeled by OAS1 and 32 P-dATP. As a comparison, the ADP-ribose isolated from PARP1 automodification reaction in the same time frame with 1 mM NAD + with a trace of 32 P-NAD + were loaded in lanes 6–8. We note that 50-fold less of the reaction were loaded in lanes 3–5 compared with lanes 6–8. (b) 15% urea-PAGE analyses of ELTA labeling reaction of ADP-ribose monomers and polymers isolated from automodification of PARP1 along with either BSA and HPF1. The first lane contained ELTA-labeling of an equal mole of 5-, 10- and 20-mer PAR. The reaction in the PARP1+BSA lane was diluted 15 times in water prior to ELTA labeling. (c) 15% urea-PAGE analyses of the ELTA labeling reaction of ADP-ribose isolated from in vitro modified ha PARP (lane 1), from untreated HaCaT cells (lane 2), from HaCaT cells treated with 1 mM H 2 O 2 for 10 min (lane 3), from HaCaT cells treated with 1 mM H 2 O 2 for 10 min, but pre-treated the cells with 20 μM PARP inhibitor Olaparib for 2 h (lane 4), or pre-treated with 1 μM PARG inhibitor PDD00017273 for 2 h (lane 5). Corresponding lysates of cells from lanes 2–5 were probed with β-actin.

Techniques Used: Polyacrylamide Gel Electrophoresis, Labeling, Isolation, In Vitro, Modification

18) Product Images from "Synthesis of [18F]fallypride in a micro-reactor"

Article Title: Synthesis of [18F]fallypride in a micro-reactor

Journal: Current radiopharmaceuticals

doi:

Influence of [ 18 F]fluoride ion/precursor solution ratio (v/v) and flow rate on RCYs of [ 18 F]fallypride at 140 °C in a 4-m micro-reactor. In these experiments, the dispensed volume of [ 18 F]F - -K + -K 2.2.2 solution was fixed at 10 μL and its
Figure Legend Snippet: Influence of [ 18 F]fluoride ion/precursor solution ratio (v/v) and flow rate on RCYs of [ 18 F]fallypride at 140 °C in a 4-m micro-reactor. In these experiments, the dispensed volume of [ 18 F]F - -K + -K 2.2.2 solution was fixed at 10 μL and its

Techniques Used: Flow Cytometry

Preparation of [ 18 F]fallypride for intravenous injection: separation and analysis. Upper panel: chromatogram of the reaction mixture obtained by infusion of  18 F - /K + -K 2.2.2 solution and precursor solution (10 μL each) into the micro-reactor at
Figure Legend Snippet: Preparation of [ 18 F]fallypride for intravenous injection: separation and analysis. Upper panel: chromatogram of the reaction mixture obtained by infusion of 18 F - /K + -K 2.2.2 solution and precursor solution (10 μL each) into the micro-reactor at

Techniques Used: Injection

19) Product Images from "CAPS Activity in Priming Vesicle Exocytosis Requires CK2 Phosphorylation *"

Article Title: CAPS Activity in Priming Vesicle Exocytosis Requires CK2 Phosphorylation *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M109.017483

CAPS is phosphorylated at N- and C-terminal Ser residues. A , MALDI-TOF spectra of CAPS phosphopeptides. Phosphopeptides enriched from CAPS tryptic peptides by Ga(III)- immobilized metal ion affinity chromatography were incubated without ( upper panel )
Figure Legend Snippet: CAPS is phosphorylated at N- and C-terminal Ser residues. A , MALDI-TOF spectra of CAPS phosphopeptides. Phosphopeptides enriched from CAPS tryptic peptides by Ga(III)- immobilized metal ion affinity chromatography were incubated without ( upper panel )

Techniques Used: Affinity Chromatography, Incubation

20) Product Images from "Tunable Open Circuit Voltage by Engineering Inorganic Cesium Lead Bromide/Iodide Perovskite Solar Cells"

Article Title: Tunable Open Circuit Voltage by Engineering Inorganic Cesium Lead Bromide/Iodide Perovskite Solar Cells

Journal: Scientific Reports

doi: 10.1038/s41598-018-20228-0

The V oc behavior of the perovskite solar cells is represented in the energy level diagrams by employing ( a ) narrow or ( b ) wide band gaps ( E g ) perovskite absorber, and ( c ) different HTMs, such as spiro -OMeTAD/Au in ( a ) and ( b ) or deeper work function (W F ) P3HT/MoO 3 /Au in ( c ). The conduction band ( E c ), valence band ( E v ), Fermi level of electrons ( E Fn ) and holes ( E Fp ) were indicated. The MoO 3 layer creates a dipole (Δ) at the interface, reducing the vacuum level energy ( E vac ). The Au performs as the electron blocking layer.
Figure Legend Snippet: The V oc behavior of the perovskite solar cells is represented in the energy level diagrams by employing ( a ) narrow or ( b ) wide band gaps ( E g ) perovskite absorber, and ( c ) different HTMs, such as spiro -OMeTAD/Au in ( a ) and ( b ) or deeper work function (W F ) P3HT/MoO 3 /Au in ( c ). The conduction band ( E c ), valence band ( E v ), Fermi level of electrons ( E Fn ) and holes ( E Fp ) were indicated. The MoO 3 layer creates a dipole (Δ) at the interface, reducing the vacuum level energy ( E vac ). The Au performs as the electron blocking layer.

Techniques Used: Blocking Assay

21) Product Images from "Simultaneous untargeted and targeted metabolomics profiling of underivatized primary metabolites in sulfur-deficient barley by ultra-high performance liquid chromatography-quadrupole/time-of-flight mass spectrometry"

Article Title: Simultaneous untargeted and targeted metabolomics profiling of underivatized primary metabolites in sulfur-deficient barley by ultra-high performance liquid chromatography-quadrupole/time-of-flight mass spectrometry

Journal: Plant Methods

doi: 10.1186/s13007-018-0329-0

Extracted ion chromatogram of amino acids and sulfur metabolites—waters acquity UPLC HSS T3 column—ESI+: 1, Proline; 2, Isoleucine; 3, Leucine; 4, Asparagine; 5, Glutamine; 6, Lysine; 7, O-Acetyl-serine; 8, Methionine; 9, Histidine; 10, Phenylalanine; 11, Arginine; 12, Tyrosine; 13, Tryptophan; 14, Thiamine; 15, Glutathione reduced; 16, S-adenosyl-methionine; 17, Glutathione oxidized
Figure Legend Snippet: Extracted ion chromatogram of amino acids and sulfur metabolites—waters acquity UPLC HSS T3 column—ESI+: 1, Proline; 2, Isoleucine; 3, Leucine; 4, Asparagine; 5, Glutamine; 6, Lysine; 7, O-Acetyl-serine; 8, Methionine; 9, Histidine; 10, Phenylalanine; 11, Arginine; 12, Tyrosine; 13, Tryptophan; 14, Thiamine; 15, Glutathione reduced; 16, S-adenosyl-methionine; 17, Glutathione oxidized

Techniques Used:

22) Product Images from "Cardioprotective effects of iron chelator HAPI and ROS-activated boronate prochelator BHAPI against catecholamine-induced oxidative cellular injury"

Article Title: Cardioprotective effects of iron chelator HAPI and ROS-activated boronate prochelator BHAPI against catecholamine-induced oxidative cellular injury

Journal: Toxicology

doi: 10.1016/j.tox.2016.10.004

Intracellular Fe-chelating efficiency of Fe chelator HAPI and prochelator BHAPI Effects of 24h-preoxidized CA isoprenaline (oxISO) or epinephrine (oxEPI) on prochelator BHAPI inside H9c2 cells were determined during 10 min with Calcein-AM assay. (A, C) Time course of BHAPI activation to effective chelator with (A) oxISO or (C) oxEPI. (B, D) Efficiency of prochelator activation to effective chelator with (B) oxISO or (D) oxEPI at time 10 min. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control, # vs. corresponding Fe chelator.
Figure Legend Snippet: Intracellular Fe-chelating efficiency of Fe chelator HAPI and prochelator BHAPI Effects of 24h-preoxidized CA isoprenaline (oxISO) or epinephrine (oxEPI) on prochelator BHAPI inside H9c2 cells were determined during 10 min with Calcein-AM assay. (A, C) Time course of BHAPI activation to effective chelator with (A) oxISO or (C) oxEPI. (B, D) Efficiency of prochelator activation to effective chelator with (B) oxISO or (D) oxEPI at time 10 min. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control, # vs. corresponding Fe chelator.

Techniques Used: Calcein AM Assay, Activation Assay

Comparison of cytotoxic effects of Fe chelator HAPI and prochelator BHAPI towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. H9c2 cells were incubated with increasing concentrations of tested compounds for 24h (A, B) or 72h (C, D) Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group.
Figure Legend Snippet: Comparison of cytotoxic effects of Fe chelator HAPI and prochelator BHAPI towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. H9c2 cells were incubated with increasing concentrations of tested compounds for 24h (A, B) or 72h (C, D) Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group.

Techniques Used: Incubation

Cellular morphology and nuclear epifluorescence staining with Hoechst 33342 and PI H9c2 cardiomyoblasts were incubated for 24 h with (A) control medium, (B) 24h-preoxidized EPI (oxEPI) alone, or in combination with studied compound (C) HAPI or (D) BHAPI, when the compound was added to oxEPI immediately before cellular experiment. Scale bars represent 100 μm.
Figure Legend Snippet: Cellular morphology and nuclear epifluorescence staining with Hoechst 33342 and PI H9c2 cardiomyoblasts were incubated for 24 h with (A) control medium, (B) 24h-preoxidized EPI (oxEPI) alone, or in combination with studied compound (C) HAPI or (D) BHAPI, when the compound was added to oxEPI immediately before cellular experiment. Scale bars represent 100 μm.

Techniques Used: Staining, Incubation

Protective effects of HAPI and BHAPI against oxEPI-induced toxicities towards isolated rat neonatal ventricularcardiomyocytes (NVCM) NVCM were incubated for 24 h with studied compounds or with their combination with 24h-preoxidized EPI (oxEPI; 700 μM). (A) Using epifluorescence microscopy, mitochondrial depolarization was assessed after loading with the JC-1 probe (red emission reflects mitochondrial inner membrane potential-dependent accumulation of probe dimers in actively respiring mitochondria, green fluorescence indicates monomers of the probe released into the cytoplasm after mitochondrial depolarization, lack of fluorescence reflects probe release from necrotic or late-stage apoptotic cells). Scale bars represent 50 μm. (B, C) Inherent toxicities of HAPI and BHAPI determined after 24 h incubation with NVCM by measurement of lactate dehydrogenase release. (D, E) Protective effects of HAPI and BHAPI against oxEPI-induced damage on NVCM assessed by measurement of lactate dehydrogenase release after 24 h incubation. Data are presented as means ± S.D.; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. oxEPI group.
Figure Legend Snippet: Protective effects of HAPI and BHAPI against oxEPI-induced toxicities towards isolated rat neonatal ventricularcardiomyocytes (NVCM) NVCM were incubated for 24 h with studied compounds or with their combination with 24h-preoxidized EPI (oxEPI; 700 μM). (A) Using epifluorescence microscopy, mitochondrial depolarization was assessed after loading with the JC-1 probe (red emission reflects mitochondrial inner membrane potential-dependent accumulation of probe dimers in actively respiring mitochondria, green fluorescence indicates monomers of the probe released into the cytoplasm after mitochondrial depolarization, lack of fluorescence reflects probe release from necrotic or late-stage apoptotic cells). Scale bars represent 50 μm. (B, C) Inherent toxicities of HAPI and BHAPI determined after 24 h incubation with NVCM by measurement of lactate dehydrogenase release. (D, E) Protective effects of HAPI and BHAPI against oxEPI-induced damage on NVCM assessed by measurement of lactate dehydrogenase release after 24 h incubation. Data are presented as means ± S.D.; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. oxEPI group.

Techniques Used: Isolation, Incubation, Epifluorescence Microscopy, Fluorescence

Process of BHAPI activation to HAPI HPLC analyses of spontaneous degradation of BHAPI and HAPI and of effects of freshly-prepared or 24h-preoxidized EPI. BHAPI or HAPI were incubated for 24 h: (A, D, G) alone, (B, E, H) with EPI or (C, F, I) with oxEPI in (A, B, C) buffered solution (pH 7.4), (D, E, F) serum-free cell-culture medium or (G, H, I) serum-free cell-culture medium with H9c2 cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. t 0 .
Figure Legend Snippet: Process of BHAPI activation to HAPI HPLC analyses of spontaneous degradation of BHAPI and HAPI and of effects of freshly-prepared or 24h-preoxidized EPI. BHAPI or HAPI were incubated for 24 h: (A, D, G) alone, (B, E, H) with EPI or (C, F, I) with oxEPI in (A, B, C) buffered solution (pH 7.4), (D, E, F) serum-free cell-culture medium or (G, H, I) serum-free cell-culture medium with H9c2 cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. t 0 .

Techniques Used: Activation Assay, High Performance Liquid Chromatography, Incubation, Cell Culture

Comparison of protective effect of HAPI and BHAPI against ISO- and EPI-induced toxicities towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. (A) HAPI or (B) BHAPI were added to freshly-prepared CA in serum-free cell-culture medium before the start of 24h cellular experiments; (C) HAPI or (D) BHAPI were added immediately before cellular experiments to CA preoxidized for 24 h in serum-free cell-culture medium; (E) HAPI or (F) BHAPI were preincubated for 24 h together with CA in serum-free cell-culture medium and then added to cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.
Figure Legend Snippet: Comparison of protective effect of HAPI and BHAPI against ISO- and EPI-induced toxicities towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. (A) HAPI or (B) BHAPI were added to freshly-prepared CA in serum-free cell-culture medium before the start of 24h cellular experiments; (C) HAPI or (D) BHAPI were added immediately before cellular experiments to CA preoxidized for 24 h in serum-free cell-culture medium; (E) HAPI or (F) BHAPI were preincubated for 24 h together with CA in serum-free cell-culture medium and then added to cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.

Techniques Used: Cell Culture

Comparison of effects of HAPI and BHAPI on cellular oxidative stress induced by 24h-preoxidized CA (oxCA – EPI and ISO) Intracellular ROS formation was determined by H 2 DCF-DA assay following the 30min treatment of H9c2 cardiomyoblasts with combination of oxCA and studied compounds. Induced intracellular fluorescence was expressed as a percentage of the group treated with oxCA alone. Cells were incubated with various concentrations of (A) HAPI or (B) BHAPI added immediately before cellular experiments to medium with oxCA (60 μM). Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.
Figure Legend Snippet: Comparison of effects of HAPI and BHAPI on cellular oxidative stress induced by 24h-preoxidized CA (oxCA – EPI and ISO) Intracellular ROS formation was determined by H 2 DCF-DA assay following the 30min treatment of H9c2 cardiomyoblasts with combination of oxCA and studied compounds. Induced intracellular fluorescence was expressed as a percentage of the group treated with oxCA alone. Cells were incubated with various concentrations of (A) HAPI or (B) BHAPI added immediately before cellular experiments to medium with oxCA (60 μM). Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.

Techniques Used: Fluorescence, Incubation

23) Product Images from "Substrate-Mediated Laser Ablation under Ambient Conditions for Spatially-Resolved Tissue Proteomics"

Article Title: Substrate-Mediated Laser Ablation under Ambient Conditions for Spatially-Resolved Tissue Proteomics

Journal: Scientific Reports

doi: 10.1038/srep18135

Evolution of the signal intensity as recorded in MALDI MS by analyzing the capture droplet after LADC experiment with the LADC laser energy/pulse from glass slide (black) versus stainless steel (red) substrates for solid BK (A), solid BK mixed with carmine (B), solid BK mixed with cytochrome C (C), and solid BK mixed with Congo red (D). Signal intensity corresponds to the averaged intensity of [M+H] + signals taken over the different measurements.
Figure Legend Snippet: Evolution of the signal intensity as recorded in MALDI MS by analyzing the capture droplet after LADC experiment with the LADC laser energy/pulse from glass slide (black) versus stainless steel (red) substrates for solid BK (A), solid BK mixed with carmine (B), solid BK mixed with cytochrome C (C), and solid BK mixed with Congo red (D). Signal intensity corresponds to the averaged intensity of [M+H] + signals taken over the different measurements.

Techniques Used: Mass Spectrometry

24) Product Images from "Analysis and Confirmation of 1,3-DMAA and 1,4-DMAA in Geranium Plants Using High Performance Liquid Chromatography with Tandem Mass Spectrometry at ng/g Concentrations"

Article Title: Analysis and Confirmation of 1,3-DMAA and 1,4-DMAA in Geranium Plants Using High Performance Liquid Chromatography with Tandem Mass Spectrometry at ng/g Concentrations

Journal: Analytical Chemistry Insights

doi: 10.4137/ACI.S10445

Typical MRM Chromatogram at 20 μg/L each for 1,3 and 1,4-DMAA analytes. Note: The retention times for the 1,3-DMAA diastereomers are 7.53 and 7.83 minutes, and 1,4-DMAA retention time is 8.17 minutes.
Figure Legend Snippet: Typical MRM Chromatogram at 20 μg/L each for 1,3 and 1,4-DMAA analytes. Note: The retention times for the 1,3-DMAA diastereomers are 7.53 and 7.83 minutes, and 1,4-DMAA retention time is 8.17 minutes.

Techniques Used:

A MRM chromatogram of Changzhou S11-1 sample. Notes: The first two peaks are 1,3-DMAA diastereomer pairs with retention times of 7.51 minutes and 7.81 minutes. The 1,4-DMAA peak retention time is 8.15 minutes. The chromatogram is produced using two mass transitions 116/99.7 m/z and 116/57 m/z.
Figure Legend Snippet: A MRM chromatogram of Changzhou S11-1 sample. Notes: The first two peaks are 1,3-DMAA diastereomer pairs with retention times of 7.51 minutes and 7.81 minutes. The 1,4-DMAA peak retention time is 8.15 minutes. The chromatogram is produced using two mass transitions 116/99.7 m/z and 116/57 m/z.

Techniques Used: Produced

A MRM chromatogram of the optimized extraction protocol for Changzhou S11–2 showing the presence of 1,3-DMAA diastereomers (peaks 1 and 2) and 1,4-DMAA (peak 3). Note: The mass transitions used are 116/99.7 m/z and 116/57 m/z.
Figure Legend Snippet: A MRM chromatogram of the optimized extraction protocol for Changzhou S11–2 showing the presence of 1,3-DMAA diastereomers (peaks 1 and 2) and 1,4-DMAA (peak 3). Note: The mass transitions used are 116/99.7 m/z and 116/57 m/z.

Techniques Used:

A typical MRM chromatogram of the Guiyang 2 sample demonstrating the absence of 1,3-DMAA and 1,4-DMAA in the geranium plant. Note: The mass transitions used are 116/99.7 m/z and 116/57 m/z.
Figure Legend Snippet: A typical MRM chromatogram of the Guiyang 2 sample demonstrating the absence of 1,3-DMAA and 1,4-DMAA in the geranium plant. Note: The mass transitions used are 116/99.7 m/z and 116/57 m/z.

Techniques Used:

A MRM chromatogram of Changzhou 3 sample showing the presence of 1,3-DMAA at a lower concentration than 1,4-DMAA. Note: Mass transitions are 116/99.7 m/z and 116/57 m/z.
Figure Legend Snippet: A MRM chromatogram of Changzhou 3 sample showing the presence of 1,3-DMAA at a lower concentration than 1,4-DMAA. Note: Mass transitions are 116/99.7 m/z and 116/57 m/z.

Techniques Used: Concentration Assay

A MRM chromatogram of the Changzhou 1 sample. Notes: The first two peaks are 1,3-DMAA diastereomer pairs with retention times of 7.51 minutes and 7.81 minutes. The 1,4-DMAA peak retention time is 8.15 minutes. The mass transitions used are 116/99.7 m/z and 116/57 m/z.
Figure Legend Snippet: A MRM chromatogram of the Changzhou 1 sample. Notes: The first two peaks are 1,3-DMAA diastereomer pairs with retention times of 7.51 minutes and 7.81 minutes. The 1,4-DMAA peak retention time is 8.15 minutes. The mass transitions used are 116/99.7 m/z and 116/57 m/z.

Techniques Used:

Chemical structures of the stereoisomers of 1,3-DMAA, 1,4-DMAA, and 2-aminoheptane with stereogenic carbons labeled (*) and their respective (R,S) configurations.
Figure Legend Snippet: Chemical structures of the stereoisomers of 1,3-DMAA, 1,4-DMAA, and 2-aminoheptane with stereogenic carbons labeled (*) and their respective (R,S) configurations.

Techniques Used: Labeling

25) Product Images from "UV laser-induced cross-linking in peptides"

Article Title: UV laser-induced cross-linking in peptides

Journal: Rapid communications in mass spectrometry : RCM

doi: 10.1002/rcm.6610

Positive-ion MALDI-TOF mass spectra of a mixture of xenopsin (M x ), interleukin (M i ) and angiotensin I (M a ) not irradiated (panel A) and irradiated for 10 sec (panel B).
Figure Legend Snippet: Positive-ion MALDI-TOF mass spectra of a mixture of xenopsin (M x ), interleukin (M i ) and angiotensin I (M a ) not irradiated (panel A) and irradiated for 10 sec (panel B).

Techniques Used: Irradiation, Size-exclusion Chromatography

26) Product Images from "Structure of the DEAH/RHA ATPase Prp43p bound to RNA implicates a pair of hairpins and motif Va in translocation along RNA"

Article Title: Structure of the DEAH/RHA ATPase Prp43p bound to RNA implicates a pair of hairpins and motif Va in translocation along RNA

Journal: RNA

doi: 10.1261/rna.060954.117

Motif Ib is not required for RNA binding or ATPase activity but is required for unwinding a 3′ overhang duplex. ( A ) Mutations in the tip of the 3′HP encompassing motif Ib (R177A or R177A/F178A) compromise unwinding of a 21-bp RNA/DNA duplex having a 27-nt 3′ RNA overhang. ( B ) Mutations in motif Ib permit RNA binding. Binding of Prp43 to the RNA unwinding substrate used in panel A was assayed by EMSA under conditions similar to the unwinding reactions but in the absence of ATP. The Prp43p–RNA complex is indicated. ( C ) Mutations in motif Ib permit ATPase activity. RNA-stimulated, Prp43-dependent ATPase activity was analyzed by TLC. The mutation E216A of motif II, the Walker B motif, was used as a negative control. ( D ) Unwinding of either a 3′ or 5′ unstructured 32-nt RNA overhang substrate with an identical 28-mer duplex. Heat denaturation of the duplex (Δ) was used as a positive control for unwinding. A line separating lanes 7 and 8 or lanes 15 and 16 indicates that a single gel image was cut and arranged to juxtapose relevant lanes.
Figure Legend Snippet: Motif Ib is not required for RNA binding or ATPase activity but is required for unwinding a 3′ overhang duplex. ( A ) Mutations in the tip of the 3′HP encompassing motif Ib (R177A or R177A/F178A) compromise unwinding of a 21-bp RNA/DNA duplex having a 27-nt 3′ RNA overhang. ( B ) Mutations in motif Ib permit RNA binding. Binding of Prp43 to the RNA unwinding substrate used in panel A was assayed by EMSA under conditions similar to the unwinding reactions but in the absence of ATP. The Prp43p–RNA complex is indicated. ( C ) Mutations in motif Ib permit ATPase activity. RNA-stimulated, Prp43-dependent ATPase activity was analyzed by TLC. The mutation E216A of motif II, the Walker B motif, was used as a negative control. ( D ) Unwinding of either a 3′ or 5′ unstructured 32-nt RNA overhang substrate with an identical 28-mer duplex. Heat denaturation of the duplex (Δ) was used as a positive control for unwinding. A line separating lanes 7 and 8 or lanes 15 and 16 indicates that a single gel image was cut and arranged to juxtapose relevant lanes.

Techniques Used: RNA Binding Assay, Activity Assay, Binding Assay, Thin Layer Chromatography, Mutagenesis, Negative Control, Positive Control

Structure of Prp43p bound to ADPNP and RNA
Figure Legend Snippet: Structure of Prp43p bound to ADPNP and RNA

Techniques Used:

The rotation of RecA2 enables RNA and ADPNP binding, and alternative motif Va conformations implicate this element as an ATP sensor. ( A , B ) The 9° rotation of RecA2 enables binding of both RNA ( A ) and ATP ( B ), implicating a mechanism by which RNA binding stimulates ATPase activity. In panel A , the movement of motifs IV and IVa in the RNA- and ADPNP-bound structure, relative to the ADP-bound structure (PDB 3KX2), is indicated by arrows. In panel B , the movement of motif VI in the RNA- and ADPNP-bound structure, relative to the ADP-bound structure, is indicated by arrows. In panels A and B , the RecA1 domain of Prp43p in the RNA-free, ADP-bound (gray) and the RNA- and ADPNP-bound (colored) states were superimposed. ( C , D ) In Prp43, as in NS3, motif Va rearranges, and movement correlates with loss of the γ-phosphate. Panel C compares the ADPNP- and RNA-bound state versus ADP-bound state of Prp43p; panel D compares the ADP–BeF 3 - and RNA-bound state versus the apo, RNA-bound state of NS3 (PDB 3O8R versus 3O8C). In each case, the RecA2 domain of the ATP-bound state (colored) was superimposed with the RecA2 domain of the contrasting states (gray). Blue highlights the static nature of motif V while red highlights the dynamic nature of motif Va. The ATP analog is shown, colored by atom type. RNA is omitted for clarity.
Figure Legend Snippet: The rotation of RecA2 enables RNA and ADPNP binding, and alternative motif Va conformations implicate this element as an ATP sensor. ( A , B ) The 9° rotation of RecA2 enables binding of both RNA ( A ) and ATP ( B ), implicating a mechanism by which RNA binding stimulates ATPase activity. In panel A , the movement of motifs IV and IVa in the RNA- and ADPNP-bound structure, relative to the ADP-bound structure (PDB 3KX2), is indicated by arrows. In panel B , the movement of motif VI in the RNA- and ADPNP-bound structure, relative to the ADP-bound structure, is indicated by arrows. In panels A and B , the RecA1 domain of Prp43p in the RNA-free, ADP-bound (gray) and the RNA- and ADPNP-bound (colored) states were superimposed. ( C , D ) In Prp43, as in NS3, motif Va rearranges, and movement correlates with loss of the γ-phosphate. Panel C compares the ADPNP- and RNA-bound state versus ADP-bound state of Prp43p; panel D compares the ADP–BeF 3 - and RNA-bound state versus the apo, RNA-bound state of NS3 (PDB 3O8R versus 3O8C). In each case, the RecA2 domain of the ATP-bound state (colored) was superimposed with the RecA2 domain of the contrasting states (gray). Blue highlights the static nature of motif V while red highlights the dynamic nature of motif Va. The ATP analog is shown, colored by atom type. RNA is omitted for clarity.

Techniques Used: Binding Assay, RNA Binding Assay, Activity Assay

27) Product Images from "Periplasmic production via the pET expression system of soluble, bioactive human growth hormone"

Article Title: Periplasmic production via the pET expression system of soluble, bioactive human growth hormone

Journal: Protein expression and purification

doi: 10.1016/j.pep.2012.11.002

Size exclusion chromatography (a–c) and MALF-TOF (d–f) analysis of TEV-TROPIN (a, d), pelB-hGH (b, e), and ompA-hGH (c, f) The retention time of the SEC calibration standards are indicated by tick marks on the x-axis. The standards include thyroglobulin 670 kDa, gamma-globulin 158 kDa, ovalbumin 44kDa, myoglobin 17 kDa, and vitamin B12 1.35 kDa. The unlabeled tick under the hGH peak in each panel corresponds to the retention time of the 17 kDa myoglobin standard. Obs: = Observed, Exp: = Expected.
Figure Legend Snippet: Size exclusion chromatography (a–c) and MALF-TOF (d–f) analysis of TEV-TROPIN (a, d), pelB-hGH (b, e), and ompA-hGH (c, f) The retention time of the SEC calibration standards are indicated by tick marks on the x-axis. The standards include thyroglobulin 670 kDa, gamma-globulin 158 kDa, ovalbumin 44kDa, myoglobin 17 kDa, and vitamin B12 1.35 kDa. The unlabeled tick under the hGH peak in each panel corresponds to the retention time of the 17 kDa myoglobin standard. Obs: = Observed, Exp: = Expected.

Techniques Used: Size-exclusion Chromatography

Reducing and non-reducing SDS-PAGE analysis of recombinant hGH Reduced TEV-TROPIN (lane 1), non-reduced TEV-TROPIN (lane 2), reduced pelB-hGH (lane 3), non-reduced pelB-hGH (lane 4), reduced ompA-hGH (lane 5), non-reduced ompA-hGH (lane 6). 7.5 μg of protein were loaded in each lane.
Figure Legend Snippet: Reducing and non-reducing SDS-PAGE analysis of recombinant hGH Reduced TEV-TROPIN (lane 1), non-reduced TEV-TROPIN (lane 2), reduced pelB-hGH (lane 3), non-reduced pelB-hGH (lane 4), reduced ompA-hGH (lane 5), non-reduced ompA-hGH (lane 6). 7.5 μg of protein were loaded in each lane.

Techniques Used: SDS Page, Recombinant

28) Product Images from "Interaction with phospholipids modulates ?-synuclein nitration and lipid-protein adduct formation"

Article Title: Interaction with phospholipids modulates ?-synuclein nitration and lipid-protein adduct formation

Journal: Biochemical Journal

doi: 10.1042/BJ20051277

Unsaturated fatty acids modulate peroxynitrite-mediated α-syn aggregation
Figure Legend Snippet: Unsaturated fatty acids modulate peroxynitrite-mediated α-syn aggregation

Techniques Used:

α-Syn binding to PC/PA liposomes modulates protein aggregation and nitration
Figure Legend Snippet: α-Syn binding to PC/PA liposomes modulates protein aggregation and nitration

Techniques Used: Binding Assay, Nitration

Formation of HNE–α-syn adducts
Figure Legend Snippet: Formation of HNE–α-syn adducts

Techniques Used:

Formation of lipid–protein adducts during peroxynitrite-mediated α-syn oxidation
Figure Legend Snippet: Formation of lipid–protein adducts during peroxynitrite-mediated α-syn oxidation

Techniques Used:

LC–MS and LC–MS/MS analyses of α-syn after tryptic digestion
Figure Legend Snippet: LC–MS and LC–MS/MS analyses of α-syn after tryptic digestion

Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

α-Syn interacts with acidic phospholipids
Figure Legend Snippet: α-Syn interacts with acidic phospholipids

Techniques Used:

Lipid–protein adduct formation as an intermediate of α-syn aggregation
Figure Legend Snippet: Lipid–protein adduct formation as an intermediate of α-syn aggregation

Techniques Used:

29) Product Images from "A Study into the ADP-Ribosylome of IFN-γ-Stimulated THP-1 Human Macrophage-like Cells Identifies ARTD8/PARP14 and ARTD9/PARP9 ADP-Ribosylation"

Article Title: A Study into the ADP-Ribosylome of IFN-γ-Stimulated THP-1 Human Macrophage-like Cells Identifies ARTD8/PARP14 and ARTD9/PARP9 ADP-Ribosylation

Journal: Journal of Proteome Research

doi: 10.1021/acs.jproteome.8b00895

Protein–protein interactions (PPI) in IFN-γ-induced ADP-ribosylome on THP-1 cells. (A) A PPI network mapping all 145 ADPr proteins from Af1521 data and visualizing their interactions (confidence interaction scores ≥700). ADPr proteins with at least one interaction with another ADPr proteins are shown. Selected GO biological processes (GO-1 to GO-7 below) from the extended list in Table S4 . (B) PPI networks of 2-fold increased or decreased ADPr proteins in the IFN-γ compared to control. (C) PPI networks from a subset of 2-fold increasing or decreasing enriched proteins using the 10H workflow in response to IFN-γ stimulation.
Figure Legend Snippet: Protein–protein interactions (PPI) in IFN-γ-induced ADP-ribosylome on THP-1 cells. (A) A PPI network mapping all 145 ADPr proteins from Af1521 data and visualizing their interactions (confidence interaction scores ≥700). ADPr proteins with at least one interaction with another ADPr proteins are shown. Selected GO biological processes (GO-1 to GO-7 below) from the extended list in Table S4 . (B) PPI networks of 2-fold increased or decreased ADPr proteins in the IFN-γ compared to control. (C) PPI networks from a subset of 2-fold increasing or decreasing enriched proteins using the 10H workflow in response to IFN-γ stimulation.

Techniques Used:

ADP-ribosylation increases during IFN-γ-induced pro-inflammatory activation of macrophages. (A) Two independent strategies, Af1521 and 10H antibody workflows, for ADP-ribosylation proteomics. (B) Antipan ADP-ribose Western blot analysis of IFN-γ-treated THP-1 cells over 24 h. (C) IFN-γ activation replicates: two sets (control or IFN-γ) of macrophage activation were used for the Af1521 workflow, and three sets were used for the 10H antibody workflow. Details about ADPr peptide data acquisition and analysis are highlighted using the second Af1521 replicate of IFN-γ-treated THP-1 cells.
Figure Legend Snippet: ADP-ribosylation increases during IFN-γ-induced pro-inflammatory activation of macrophages. (A) Two independent strategies, Af1521 and 10H antibody workflows, for ADP-ribosylation proteomics. (B) Antipan ADP-ribose Western blot analysis of IFN-γ-treated THP-1 cells over 24 h. (C) IFN-γ activation replicates: two sets (control or IFN-γ) of macrophage activation were used for the Af1521 workflow, and three sets were used for the 10H antibody workflow. Details about ADPr peptide data acquisition and analysis are highlighted using the second Af1521 replicate of IFN-γ-treated THP-1 cells.

Techniques Used: Activation Assay, Western Blot

IFN-γ increased ARTD8/PARP14 and ARTD9/PARP9 ADP-ribosylation. (A) A pan ADP-ribose Western blot analysis of control and IFN-γ-treated THP-1 cells after 10H IP or incubation with IgG. (B) A comparison of enriched proteins using a Venn diagram between 10H and IgG. The plot showing the log 2 (abundance ratio (10H/IgG)) and −log( p -value) of 551 shared proteins (Venn diagram). 114 proteins passed the threshold of abundance ratio (10H/IgG) > 10-fold and p -value
Figure Legend Snippet: IFN-γ increased ARTD8/PARP14 and ARTD9/PARP9 ADP-ribosylation. (A) A pan ADP-ribose Western blot analysis of control and IFN-γ-treated THP-1 cells after 10H IP or incubation with IgG. (B) A comparison of enriched proteins using a Venn diagram between 10H and IgG. The plot showing the log 2 (abundance ratio (10H/IgG)) and −log( p -value) of 551 shared proteins (Venn diagram). 114 proteins passed the threshold of abundance ratio (10H/IgG) > 10-fold and p -value

Techniques Used: Western Blot, Incubation

30) Product Images from "High-resolution mass spectrometry analysis of protein oxidations and resultant loss of function"

Article Title: High-resolution mass spectrometry analysis of protein oxidations and resultant loss of function

Journal: Biochemical Society transactions

doi: 10.1042/BST0361037

LTQ tandem mass spectrum of the triply oxidized αB-crystallin peptide 57 APSWFDTGLSEMR 69 . The b 4 ion ( m/z 474.2) is 218.1 bigger than the b 3 ion ( m/z 256.1), showing that the Trp residue contains two oxygen atoms (186.1 + 2 × 16). Similarly,
Figure Legend Snippet: LTQ tandem mass spectrum of the triply oxidized αB-crystallin peptide 57 APSWFDTGLSEMR 69 . The b 4 ion ( m/z 474.2) is 218.1 bigger than the b 3 ion ( m/z 256.1), showing that the Trp residue contains two oxygen atoms (186.1 + 2 × 16). Similarly,

Techniques Used:

31) Product Images from "The S. pombe “cytokinesis” NDR kinase Sid2 activates Fin1 NIMA kinase to control mitotic commitment via Pom1/Wee1"

Article Title: The S. pombe “cytokinesis” NDR kinase Sid2 activates Fin1 NIMA kinase to control mitotic commitment via Pom1/Wee1

Journal: Nature cell biology

doi: 10.1038/ncb2514

Sid2 phosphorylation of Fin1 on serines 377, 526 and 698 promotes Fin1 activity in G2 phase before a peak of each kinase activity accompanies mitotic progression (a, b) Fin1 Kinase assays from size selected cultures in which His tagged Fin1.KD was labeled with 32 PγATP to quantitate activity that is plotted alongside the septation profile. (a) wild type (b) Small G2 sid2.as4 skp1.A4 cells were isolated from a culture grown at 25°C and immediately shifted to 36°C to inactivate Skp1 (and so preserve activated Fin1) at t=0. The culture was split in two and 20 μM 3-MB-PP1 (left assay) or solvent alone (right assay) added after the first division at t = 180. (c, d) Fin1 immunoprecipitates from asynchronous skp1.A4 (c) or cell size selected sid2.as4 skp1.A4 (d) cultures were split in two and probed with antibodies that recognise the indicated phosphorylation sites or polyclonal antibodies that recognise the non-catalytic domain of Fin1. See Supplementary Figure 3g for details of the scheme used for each of the three identical cultures used to generate the samples and Supplmentary Figure 3h for the phenotypic characterisation of one of the three cultures. Samples from asynchronous cultures are run in the left lane in each case to provide a reference standard. (e) Sid2 immunoprecipitates were isolated from asynchronous cultures and employed in in vitro kinase assays utilising 32 PγATP and either recombinant Fin1.KD or casein as indicated. Plots show activity per unit protein (i.e. specific activity) f) Blots with the indicated antibodies of in vitro kinase assays in which the indicated forms of Sid2 were isolated from the respective strains and combined with recombinant Fin1.KDnHis. (g) The incorporation of 32 P into casein from 32 PγATP was used to monitor Sid2 activity in size selected wild type cultures. (h) Sid2 immunoprecipitates were processed as for panel g with the exception that the shorter Fin1.FP1 (non-catalytic C terminal domain 21 ) was used as a substrate and the F1S698P antibody was used to develop the assay with the secondary reagent BCIP. The loading of the Sid2.250 36°C sample in the second to last lane was four times that in other lanes to ensure that the basal level dictated by the reduced level of Sid2.250 protein in the 36°C sample was representative of the reference point for normalisation.
Figure Legend Snippet: Sid2 phosphorylation of Fin1 on serines 377, 526 and 698 promotes Fin1 activity in G2 phase before a peak of each kinase activity accompanies mitotic progression (a, b) Fin1 Kinase assays from size selected cultures in which His tagged Fin1.KD was labeled with 32 PγATP to quantitate activity that is plotted alongside the septation profile. (a) wild type (b) Small G2 sid2.as4 skp1.A4 cells were isolated from a culture grown at 25°C and immediately shifted to 36°C to inactivate Skp1 (and so preserve activated Fin1) at t=0. The culture was split in two and 20 μM 3-MB-PP1 (left assay) or solvent alone (right assay) added after the first division at t = 180. (c, d) Fin1 immunoprecipitates from asynchronous skp1.A4 (c) or cell size selected sid2.as4 skp1.A4 (d) cultures were split in two and probed with antibodies that recognise the indicated phosphorylation sites or polyclonal antibodies that recognise the non-catalytic domain of Fin1. See Supplementary Figure 3g for details of the scheme used for each of the three identical cultures used to generate the samples and Supplmentary Figure 3h for the phenotypic characterisation of one of the three cultures. Samples from asynchronous cultures are run in the left lane in each case to provide a reference standard. (e) Sid2 immunoprecipitates were isolated from asynchronous cultures and employed in in vitro kinase assays utilising 32 PγATP and either recombinant Fin1.KD or casein as indicated. Plots show activity per unit protein (i.e. specific activity) f) Blots with the indicated antibodies of in vitro kinase assays in which the indicated forms of Sid2 were isolated from the respective strains and combined with recombinant Fin1.KDnHis. (g) The incorporation of 32 P into casein from 32 PγATP was used to monitor Sid2 activity in size selected wild type cultures. (h) Sid2 immunoprecipitates were processed as for panel g with the exception that the shorter Fin1.FP1 (non-catalytic C terminal domain 21 ) was used as a substrate and the F1S698P antibody was used to develop the assay with the secondary reagent BCIP. The loading of the Sid2.250 36°C sample in the second to last lane was four times that in other lanes to ensure that the basal level dictated by the reduced level of Sid2.250 protein in the 36°C sample was representative of the reference point for normalisation.

Techniques Used: Activity Assay, Labeling, Isolation, In Vitro, Recombinant

Fin1 kinase is destroyed twice each cell cycle in a Cullin, Fin1 and Sid2 dependent manner (a, b, d, e, h-j) Fin1 levels were normalised to those of Cdc2 kinase in the same lane on the same blot and plotted against time as cells transit the cell cycle (for images of blots see Supplementary Figure 1b ). (a) Fin1 levels declined at two points in wild type cultures; mid-G2 (grey arrow “G2”) and during septation (open arrow “C”). Destruction was seen irrespective of whether the culture was maintained at 25°C throughout the experiment, or shifted to 36°C immediately after size selection ( Supplementary Figure 4c ). (b) Oscillations in Fin1 levels were not seen after synchronised skp1.A4 cultures were shifted to 36°C immediately after size selection at 25°C to inactivate Skp1. (c, g) Normalised Fin1 levels in blots of asynchronous or cdc25.22 arrested double mutant cultures reveal three fold increases in Fin1 levels in the fin1.K33RN165A “kinase dead” and sid2.250 backgrounds. (d) Fin1 levels did not fluctuate as fin1.K33RN165A cultures transited a synchronised cell cycle. (e) Strikingly the levels of both the inactive fin1.K33RN165A protein and the GFP tagged wild type protein oscillate as cells transit the cell cycle when a wild type Fin1.GFP fusion protein was constitutively expressed within the same cells. (f) Fin1 immunoprecipitates from asynchronous cells were employed in kinase assays that used recombinant or casein as substrates. (g) Left: 210 and 240 mins refers to the duration of incubation at 36°C to inactivate and arrest cell cycle progression at the G2/M boundary. Right: FACS profiles of DNA content demonstrate G2 arrest in all strains. (h-j) Assessing the impact of Sid2/Mob1 function upon Fin1 levels in size selected synchronised cultures. (h, j) sid2.250 and mob1.E9 cultures were maintained at 25°C during transit through the first cell division before a portion of the culture was shifted to 36°C to inactivate the kinase/regulatory subunit. (i) A sid2.as4 culture was split into three after the first wave of septation was complete ( Supplementary Figure 1c ) and either nothing, methanol or 3-MB-PP1 in methanol were added to a final concentration of 20μM at time point 190.
Figure Legend Snippet: Fin1 kinase is destroyed twice each cell cycle in a Cullin, Fin1 and Sid2 dependent manner (a, b, d, e, h-j) Fin1 levels were normalised to those of Cdc2 kinase in the same lane on the same blot and plotted against time as cells transit the cell cycle (for images of blots see Supplementary Figure 1b ). (a) Fin1 levels declined at two points in wild type cultures; mid-G2 (grey arrow “G2”) and during septation (open arrow “C”). Destruction was seen irrespective of whether the culture was maintained at 25°C throughout the experiment, or shifted to 36°C immediately after size selection ( Supplementary Figure 4c ). (b) Oscillations in Fin1 levels were not seen after synchronised skp1.A4 cultures were shifted to 36°C immediately after size selection at 25°C to inactivate Skp1. (c, g) Normalised Fin1 levels in blots of asynchronous or cdc25.22 arrested double mutant cultures reveal three fold increases in Fin1 levels in the fin1.K33RN165A “kinase dead” and sid2.250 backgrounds. (d) Fin1 levels did not fluctuate as fin1.K33RN165A cultures transited a synchronised cell cycle. (e) Strikingly the levels of both the inactive fin1.K33RN165A protein and the GFP tagged wild type protein oscillate as cells transit the cell cycle when a wild type Fin1.GFP fusion protein was constitutively expressed within the same cells. (f) Fin1 immunoprecipitates from asynchronous cells were employed in kinase assays that used recombinant or casein as substrates. (g) Left: 210 and 240 mins refers to the duration of incubation at 36°C to inactivate and arrest cell cycle progression at the G2/M boundary. Right: FACS profiles of DNA content demonstrate G2 arrest in all strains. (h-j) Assessing the impact of Sid2/Mob1 function upon Fin1 levels in size selected synchronised cultures. (h, j) sid2.250 and mob1.E9 cultures were maintained at 25°C during transit through the first cell division before a portion of the culture was shifted to 36°C to inactivate the kinase/regulatory subunit. (i) A sid2.as4 culture was split into three after the first wave of septation was complete ( Supplementary Figure 1c ) and either nothing, methanol or 3-MB-PP1 in methanol were added to a final concentration of 20μM at time point 190.

Techniques Used: Selection, Mutagenesis, Recombinant, Incubation, FACS, Concentration Assay

Inhibition of Sid2 or Fin1 delays mitotic commitment (a) 3-MB-PP1 was added to asynchronous cultures of wild type, fin1.as3 and sid2.as4 cells and the mitotic index monitored by anti-α-tubulin immunofluorescence at the indicated times. The analogue transiently inhibited mitotic commitment of fin1.as3 and sid2.as4 but not wild type cells. (b-d) Wild type, fin1.as3 and sid2.as4 cultures were synchronised with respect to cell cycle progression by size selection and split into three equal cultures after the first round of septation. Methanol (MeOH), or 3-MB-PP1 (to a final concentration of 20 μM) in MeOH were added to two of these sub-cultures at 160 minutes. Commitment to mitosis was monitored by the spindle index or phospho-histone H3 reactivity, as indicated. Addition of solvent alone had no impact upon cell cycle progression while addition of analogue in solvent delayed mitotic commitment in fin1.as3 and sid2.as4 but had no impact upon wild type cells.
Figure Legend Snippet: Inhibition of Sid2 or Fin1 delays mitotic commitment (a) 3-MB-PP1 was added to asynchronous cultures of wild type, fin1.as3 and sid2.as4 cells and the mitotic index monitored by anti-α-tubulin immunofluorescence at the indicated times. The analogue transiently inhibited mitotic commitment of fin1.as3 and sid2.as4 but not wild type cells. (b-d) Wild type, fin1.as3 and sid2.as4 cultures were synchronised with respect to cell cycle progression by size selection and split into three equal cultures after the first round of septation. Methanol (MeOH), or 3-MB-PP1 (to a final concentration of 20 μM) in MeOH were added to two of these sub-cultures at 160 minutes. Commitment to mitosis was monitored by the spindle index or phospho-histone H3 reactivity, as indicated. Addition of solvent alone had no impact upon cell cycle progression while addition of analogue in solvent delayed mitotic commitment in fin1.as3 and sid2.as4 but had no impact upon wild type cells.

Techniques Used: Inhibition, Immunofluorescence, Selection, Concentration Assay

Sid2 and Fin1 target Cdr1/Cdr2/Pom1 to control mitotic commitment via Wee1 (a) Cartoon detailing the analogue washout approach. (b-f) 5 hours after the addition of 3-MB-PP1 to early log phase cultures of the indicated strains cells were filtered from the culture and re-suspended at the same density in growth medium that contained no inhibitor. Restoration of Sid2 and Fin1 function induced a burst of mitosis. Importantly, restoration of Sid2 function failed to induce mitotic commitment when Fin1 kinase was inactivated by the fin1.KD mutations (b). (c) Restoring Sid2 activity in strains harbouring mutation of the candidate Sid2 phosphorylation sites in Fin1 suggests that Sid2 can activate Fin1 by phosphorylating the serine at either 377, 526 or 698. Both Sid2 and Fin1 were able to induce mitosis when the cdc2.3w mutation compromised sensitivity to Cdc25, but not when the cdc2.1w mutation compromised Wee1 inhibition of Cdc2 (d), or the functions of Pom1, Cdr1 or Cdr2 are ablated (e, f). (g) Pom1.GFP signals in the indicated strain backgrounds. h) A cartoon depicting the model for G2/M control by Sid2/Fin1. P represents phosphorylation, while Ub represents Ubiquitin conjugation. Fin1 is activated in G2 by phosphorylation by Sid2. This promotes mitotic commitment via modulation of the Geometry network, however, the exact mechanism remains to be determined. Activated Fin1 promotes its own destruction, thereby limiting its activity temporally. Our current lack of understanding of the means by which Fin1 regulates the Pom1/Cdr1/Cdr2/Wee1 cell Geometry Network is represented by incorporating all members of this pathway that are required for Fin1 to regulate mitotic commitment within a single box. The question mark to the left of this box reflects our lack of knowledge as to whether it is the Sid2 or the auto-phosphorylated form of Fin1 that is responsible for the control of the Cell Geometry Network. The question mark beneath Sid1/Cdc14 ref lects our ignorance as to the nature of the cue in G2 phase that triggers this pathway. See text for further details.
Figure Legend Snippet: Sid2 and Fin1 target Cdr1/Cdr2/Pom1 to control mitotic commitment via Wee1 (a) Cartoon detailing the analogue washout approach. (b-f) 5 hours after the addition of 3-MB-PP1 to early log phase cultures of the indicated strains cells were filtered from the culture and re-suspended at the same density in growth medium that contained no inhibitor. Restoration of Sid2 and Fin1 function induced a burst of mitosis. Importantly, restoration of Sid2 function failed to induce mitotic commitment when Fin1 kinase was inactivated by the fin1.KD mutations (b). (c) Restoring Sid2 activity in strains harbouring mutation of the candidate Sid2 phosphorylation sites in Fin1 suggests that Sid2 can activate Fin1 by phosphorylating the serine at either 377, 526 or 698. Both Sid2 and Fin1 were able to induce mitosis when the cdc2.3w mutation compromised sensitivity to Cdc25, but not when the cdc2.1w mutation compromised Wee1 inhibition of Cdc2 (d), or the functions of Pom1, Cdr1 or Cdr2 are ablated (e, f). (g) Pom1.GFP signals in the indicated strain backgrounds. h) A cartoon depicting the model for G2/M control by Sid2/Fin1. P represents phosphorylation, while Ub represents Ubiquitin conjugation. Fin1 is activated in G2 by phosphorylation by Sid2. This promotes mitotic commitment via modulation of the Geometry network, however, the exact mechanism remains to be determined. Activated Fin1 promotes its own destruction, thereby limiting its activity temporally. Our current lack of understanding of the means by which Fin1 regulates the Pom1/Cdr1/Cdr2/Wee1 cell Geometry Network is represented by incorporating all members of this pathway that are required for Fin1 to regulate mitotic commitment within a single box. The question mark to the left of this box reflects our lack of knowledge as to whether it is the Sid2 or the auto-phosphorylated form of Fin1 that is responsible for the control of the Cell Geometry Network. The question mark beneath Sid1/Cdc14 ref lects our ignorance as to the nature of the cue in G2 phase that triggers this pathway. See text for further details.

Techniques Used: Activity Assay, Mutagenesis, Inhibition, Conjugation Assay

32) Product Images from "Identification of Clostridium spp. derived from a sheep and cattle slaughterhouse by matrix-assisted laser desorption and ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rDNA sequencing"

Article Title: Identification of Clostridium spp. derived from a sheep and cattle slaughterhouse by matrix-assisted laser desorption and ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rDNA sequencing

Journal: Journal of Food Science and Technology

doi: 10.1007/s13197-018-3255-2

Comparing direct transfer and extended direct transfer sample preparation methods for identification of Clostridium species by MALDI-TOF MS at different days of anaerobic incubation (1, 3, and 5). Each isolate was tested three times for each method of sample preparation at each day
Figure Legend Snippet: Comparing direct transfer and extended direct transfer sample preparation methods for identification of Clostridium species by MALDI-TOF MS at different days of anaerobic incubation (1, 3, and 5). Each isolate was tested three times for each method of sample preparation at each day

Techniques Used: Sample Prep, Mass Spectrometry, Incubation

Overview of Bruker MALDI-TOF MS identification of Clostridium isolates at the species level. MALDI-TOF MS identified 94% isolates while 6% of isolates were not identified by MALDI-TOF MS. These isolates were identified via 16S rDNA as C. estertheticum (n = 8), C. frigidicarnis (n = 5), and C. gasigenes (n = 3) species
Figure Legend Snippet: Overview of Bruker MALDI-TOF MS identification of Clostridium isolates at the species level. MALDI-TOF MS identified 94% isolates while 6% of isolates were not identified by MALDI-TOF MS. These isolates were identified via 16S rDNA as C. estertheticum (n = 8), C. frigidicarnis (n = 5), and C. gasigenes (n = 3) species

Techniques Used: Mass Spectrometry

33) Product Images from "The Oligomerization Domain of VP3, the Scaffolding Protein of Infectious Bursal Disease Virus, Plays a Critical Role in Capsid Assembly"

Article Title: The Oligomerization Domain of VP3, the Scaffolding Protein of Infectious Bursal Disease Virus, Plays a Critical Role in Capsid Assembly

Journal: Journal of Virology

doi: 10.1128/JVI.77.11.6438-6449.2003

Effect of VP1 coexpression on His-VP3 proteolytic cleavage. (A) Detection of VP3-VP1 complexes. H5 cells were infected with FB/His-VP3 or FB/His-VP3-VP1. At 72 h postinfection, cells were harvested, and the corresponding extracts were subjected to IMAC purification. Samples corresponding to total cell extracts (T) and IMAC-purified polypeptides (P) were analyzed by SDS-PAGE. After electrophoresis, gels were fixed and silver stained. The positions of molecular mass markers are indicated (in kilodaltons). (B) Western blot analysis of extracts from H5 cells infected with FB/His-VP3 or FB/His-VP3-VP1 or coinfected with FB/His-VP3 and FB/His-VP1. Infected cells were harvested at 72 h postinfection, resuspended in lysis buffer, and subjected to IMAC purification. Samples were subjected to SDS-PAGE and Western blot analysis with rabbit anti-VP3 serum, followed by addition of horseradish peroxidase-conjugated goat anti-rat immunoglobulin. The signal was detected by enhanced chemiluminescence. The positions of molecular mass markers (in kilodaltons) are indicated.
Figure Legend Snippet: Effect of VP1 coexpression on His-VP3 proteolytic cleavage. (A) Detection of VP3-VP1 complexes. H5 cells were infected with FB/His-VP3 or FB/His-VP3-VP1. At 72 h postinfection, cells were harvested, and the corresponding extracts were subjected to IMAC purification. Samples corresponding to total cell extracts (T) and IMAC-purified polypeptides (P) were analyzed by SDS-PAGE. After electrophoresis, gels were fixed and silver stained. The positions of molecular mass markers are indicated (in kilodaltons). (B) Western blot analysis of extracts from H5 cells infected with FB/His-VP3 or FB/His-VP3-VP1 or coinfected with FB/His-VP3 and FB/His-VP1. Infected cells were harvested at 72 h postinfection, resuspended in lysis buffer, and subjected to IMAC purification. Samples were subjected to SDS-PAGE and Western blot analysis with rabbit anti-VP3 serum, followed by addition of horseradish peroxidase-conjugated goat anti-rat immunoglobulin. The signal was detected by enhanced chemiluminescence. The positions of molecular mass markers (in kilodaltons) are indicated.

Techniques Used: Infection, Purification, SDS Page, Electrophoresis, Staining, Western Blot, Lysis

Mapping VP3 proteolytic cleavage site. (A) The diagram shows the set of VP3 C-terminal deletion mutants used for mapping the VP3 cleavage site. (B) Western blot analysis of IMAC-purified polypeptides encoded by the different VP3 C-terminal deletion mutants. Extracts from rBV-infected cells were subjected to IMAC. Affinity-purified polypeptides were subjected to SDS-PAGE and Western blot analysis with rabbit anti-VP3 serum, followed by addition of horseradish peroxidase-conjugated goat anti-rat immunoglobulin. The signal was detected by enhanced chemiluminescence. The positions of molecular mass markers are shown (in kilodaltons). Arrows indicate the position corresponding to full-length (F) and C-terminally trimmed (T) His-VP3 polypeptide products.
Figure Legend Snippet: Mapping VP3 proteolytic cleavage site. (A) The diagram shows the set of VP3 C-terminal deletion mutants used for mapping the VP3 cleavage site. (B) Western blot analysis of IMAC-purified polypeptides encoded by the different VP3 C-terminal deletion mutants. Extracts from rBV-infected cells were subjected to IMAC. Affinity-purified polypeptides were subjected to SDS-PAGE and Western blot analysis with rabbit anti-VP3 serum, followed by addition of horseradish peroxidase-conjugated goat anti-rat immunoglobulin. The signal was detected by enhanced chemiluminescence. The positions of molecular mass markers are shown (in kilodaltons). Arrows indicate the position corresponding to full-length (F) and C-terminally trimmed (T) His-VP3 polypeptide products.

Techniques Used: Western Blot, Purification, Infection, Affinity Purification, SDS Page

34) Product Images from "Icariin Metabolism by Human Intestinal Microflora"

Article Title: Icariin Metabolism by Human Intestinal Microflora

Journal: Molecules

doi: 10.3390/molecules21091158

MS spectra of icariin and its metabolites. Thermo Fisher Scientific LCQ fleet instrument (Thermo Scientific, Waltham, MA, USA) was used for electrospray ionization mass spectrometry (ESI-MS) analysis. ESI condition: spray voltage, 5.4 kV; sheath gas, 15 arbitrary units; auxiliary gas, five arbitrary units; heated capillary temperature, 275 °C; capillary voltage, 27 V; and tube lens, 100 V.
Figure Legend Snippet: MS spectra of icariin and its metabolites. Thermo Fisher Scientific LCQ fleet instrument (Thermo Scientific, Waltham, MA, USA) was used for electrospray ionization mass spectrometry (ESI-MS) analysis. ESI condition: spray voltage, 5.4 kV; sheath gas, 15 arbitrary units; auxiliary gas, five arbitrary units; heated capillary temperature, 275 °C; capillary voltage, 27 V; and tube lens, 100 V.

Techniques Used: Mass Spectrometry

Proposed metabolic pathway of icariin in human intestine.
Figure Legend Snippet: Proposed metabolic pathway of icariin in human intestine.

Techniques Used:

Time-dependent biotransformation of icariin by ( a ) Streptococcus sp. MRG-ICA-B and ( b ) Blautia sp. MRG-PMF-1.
Figure Legend Snippet: Time-dependent biotransformation of icariin by ( a ) Streptococcus sp. MRG-ICA-B and ( b ) Blautia sp. MRG-PMF-1.

Techniques Used: Peptide Mass Fingerprinting

HPLC chromatogram changes at 270 nm absorption over icariin metabolism by Blautia sp. MRG-PMF1.
Figure Legend Snippet: HPLC chromatogram changes at 270 nm absorption over icariin metabolism by Blautia sp. MRG-PMF1.

Techniques Used: High Performance Liquid Chromatography

HPLC chromatograms of icariin biotransformation products. Each chromatogram was obtained from the different human intestinal microflora. Microflora a , b and c showed icariside II, icaritin and desmethylicaritin formation, respectively, after 48 h.
Figure Legend Snippet: HPLC chromatograms of icariin biotransformation products. Each chromatogram was obtained from the different human intestinal microflora. Microflora a , b and c showed icariside II, icaritin and desmethylicaritin formation, respectively, after 48 h.

Techniques Used: High Performance Liquid Chromatography

35) Product Images from "Multiwalled Carbon Nanotube for One-Step Cleanup of 21 Mycotoxins in Corn and Wheat Prior to Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry Analysis"

Article Title: Multiwalled Carbon Nanotube for One-Step Cleanup of 21 Mycotoxins in Corn and Wheat Prior to Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry Analysis

Journal: Toxins

doi: 10.3390/toxins10100409

Effects of three MWCNT sorbents on recovery (%) of mycotoxins in corn. MWCNT, multiwalled carbon nanotube; MWCNT–COOH, carboxylic MWCNT; MWCNT–OH, hydroxyl MWCNT; DON, deoxynivalenol; NIV, nivalenol; AFB 1 , AFB 2 , AFG 1 , AFG 2 , aflatoxins; 15-AcDON, 15-acetyldeoxynivalenol; 3-AcDON, 3-acetyldeoxynivalenol; FUS-X, fusarenon X; DAS, diacetoxyscirpenol; OTA, ochratoxin A; OTB, ochratoxin B; T-2, T-2 toxin; HT-2, HT-2 toxin; NEO, neosolaniol; ZEN, zearalenone; α-ZOL, α-zearalenol; β-ZOL, β-zearalenol; ZAN, zearalanone; α-ZAL, α-zearalanol; β-ZAL, β-zearalanol. Vertical bar represents ± standard error ( n = 3).
Figure Legend Snippet: Effects of three MWCNT sorbents on recovery (%) of mycotoxins in corn. MWCNT, multiwalled carbon nanotube; MWCNT–COOH, carboxylic MWCNT; MWCNT–OH, hydroxyl MWCNT; DON, deoxynivalenol; NIV, nivalenol; AFB 1 , AFB 2 , AFG 1 , AFG 2 , aflatoxins; 15-AcDON, 15-acetyldeoxynivalenol; 3-AcDON, 3-acetyldeoxynivalenol; FUS-X, fusarenon X; DAS, diacetoxyscirpenol; OTA, ochratoxin A; OTB, ochratoxin B; T-2, T-2 toxin; HT-2, HT-2 toxin; NEO, neosolaniol; ZEN, zearalenone; α-ZOL, α-zearalenol; β-ZOL, β-zearalenol; ZAN, zearalanone; α-ZAL, α-zearalanol; β-ZAL, β-zearalanol. Vertical bar represents ± standard error ( n = 3).

Techniques Used: Ziehl-Neelsen Stain

36) Product Images from "Cysteine residues contribute to the dimerization and enzymatic activity of human nuclear dUTP nucleotidohydrolase (nDut)"

Article Title: Cysteine residues contribute to the dimerization and enzymatic activity of human nuclear dUTP nucleotidohydrolase (nDut)

Journal: Protein Science : A Publication of the Protein Society

doi: 10.1002/pro.3481

Cysteine 3 is a critical residue in stabilizing the secondary structure of nuclear dUTPase. The expression of nDut.Ctag, nDut.Ntag and cysteine to alanine mutants in U‐2 OS cells implicate C3 as a critical residue in stabilizing the higher order structure of n.Dut. Two constructs of each set of proteins contained either a hexahistidine C‐terminus tag (A, C) or hexahistidine N‐terminus tag (B, D). 3 μg of each plasmid were transiently transfected into U2‐OS cells followed by a 24‐h incubation. The cells were harvested and 20 μg of total cell extract was applied to a 4–20% tris‐glycine SDS‐PAGE (±) BME as indicated. Western blot analysis was then preformed using a histidine primary antibody. The predicted molecular weight of nDut.Ntag is 21,188 Da. This includes the existence of a 28 amino acid leader sequence (3458 Da). The nDut.Ctag construct is void of this leader sequence and is predicted to be 18,571 Da. The lower molecular weight band observed in (A) and (C) is likely a truncated form of dUTPase (M24) (Fig. S3 ).
Figure Legend Snippet: Cysteine 3 is a critical residue in stabilizing the secondary structure of nuclear dUTPase. The expression of nDut.Ctag, nDut.Ntag and cysteine to alanine mutants in U‐2 OS cells implicate C3 as a critical residue in stabilizing the higher order structure of n.Dut. Two constructs of each set of proteins contained either a hexahistidine C‐terminus tag (A, C) or hexahistidine N‐terminus tag (B, D). 3 μg of each plasmid were transiently transfected into U2‐OS cells followed by a 24‐h incubation. The cells were harvested and 20 μg of total cell extract was applied to a 4–20% tris‐glycine SDS‐PAGE (±) BME as indicated. Western blot analysis was then preformed using a histidine primary antibody. The predicted molecular weight of nDut.Ntag is 21,188 Da. This includes the existence of a 28 amino acid leader sequence (3458 Da). The nDut.Ctag construct is void of this leader sequence and is predicted to be 18,571 Da. The lower molecular weight band observed in (A) and (C) is likely a truncated form of dUTPase (M24) (Fig. S3 ).

Techniques Used: Expressing, Construct, Plasmid Preparation, Transfection, Incubation, SDS Page, Western Blot, Molecular Weight, Sequencing

An intermolecular disulfide bridge formation between two cysteine 3 residues is essential for nuclear dUTPase dimer formation. A 12% Tris‐Glycine SDS‐PAGE coomassie stained gel of recombinant Wt nDut.Ntag and cysteine to alanine mutants in (A) non‐reducing (‐BME) or (B) reducing conditions (+BME). (C) A 16% Tris‐Glycine SDS‐PAGE coomassie stained gel of recombinant Wt nDut.Ctag (+/‐) BME. Predicted molecular weights and migration behavior on SDS‐PAGE is reflective of what is seen in Figure 2 . Mass spectrometry was performed with 2 μg of both the full length Wt nDut.ntag (D) and the C3A mutant (E). The peak of interest corresponds to the dimeric (42 KDa) state of the protein which can be visualized in the Wt (D) spectrum. As shown, this peak is significantly diminished in the C3A spectrum.
Figure Legend Snippet: An intermolecular disulfide bridge formation between two cysteine 3 residues is essential for nuclear dUTPase dimer formation. A 12% Tris‐Glycine SDS‐PAGE coomassie stained gel of recombinant Wt nDut.Ntag and cysteine to alanine mutants in (A) non‐reducing (‐BME) or (B) reducing conditions (+BME). (C) A 16% Tris‐Glycine SDS‐PAGE coomassie stained gel of recombinant Wt nDut.Ctag (+/‐) BME. Predicted molecular weights and migration behavior on SDS‐PAGE is reflective of what is seen in Figure 2 . Mass spectrometry was performed with 2 μg of both the full length Wt nDut.ntag (D) and the C3A mutant (E). The peak of interest corresponds to the dimeric (42 KDa) state of the protein which can be visualized in the Wt (D) spectrum. As shown, this peak is significantly diminished in the C3A spectrum.

Techniques Used: SDS Page, Staining, Recombinant, Migration, Mass Spectrometry, Mutagenesis

37) Product Images from "Role of Lipid Peroxidation and PPAR-? in Amplifying Glucose-Stimulated Insulin Secretion"

Article Title: Role of Lipid Peroxidation and PPAR-? in Amplifying Glucose-Stimulated Insulin Secretion

Journal: Diabetes

doi: 10.2337/db11-0347

NAC reduces ROS production, 4-HNE generation, and insulin secretion in β-cells. A : INS-1E cells were incubated for 16 h with the indicated glucose levels without or with 1 mmol/L NAC. ROS production was determined with the carboxy-DCF-fluorescence assay. Glc, glucose. * P
Figure Legend Snippet: NAC reduces ROS production, 4-HNE generation, and insulin secretion in β-cells. A : INS-1E cells were incubated for 16 h with the indicated glucose levels without or with 1 mmol/L NAC. ROS production was determined with the carboxy-DCF-fluorescence assay. Glc, glucose. * P

Techniques Used: Incubation, Fluorescence, Gas Chromatography

PPAR-δ mediates 4-HNE effects in β-cells. A : PPAR-δ expression was silenced in INS-1E cells with targeted siRNA sequences (Si), as described in research design and methods . The cells were then incubated for 24 h at the indicated glucose concentrations, washed, processed, and taken for the standard GSIS analysis, as described in the legend to Fig. 3 A . Cells transfected with scrambled RNA (Sc) served as controls. C, nontransfected cells; Glc, glucose. * P
Figure Legend Snippet: PPAR-δ mediates 4-HNE effects in β-cells. A : PPAR-δ expression was silenced in INS-1E cells with targeted siRNA sequences (Si), as described in research design and methods . The cells were then incubated for 24 h at the indicated glucose concentrations, washed, processed, and taken for the standard GSIS analysis, as described in the legend to Fig. 3 A . Cells transfected with scrambled RNA (Sc) served as controls. C, nontransfected cells; Glc, glucose. * P

Techniques Used: Expressing, Incubation, Transfection, Gas Chromatography

Effect of high glucose on PUFA peroxidation in β-cells. A : INS-1E cells were exposed to the indicated d -glucose (D-Glc) and l -glucose (L-Glc) concentrations for 48 h; during the last 16 h, the cells were incubated with serum-free culture medium with the same additions. The media (10 mL) were then collected, extracted, and analyzed by HPLC. Data are given as nanogram 4-HNE per milligram cellular protein. Representative HPLC tracings are depicted ( inset ) and the arrows point to 4-HNE peaks. Results are mean ± SEM, n = 3–4. * P
Figure Legend Snippet: Effect of high glucose on PUFA peroxidation in β-cells. A : INS-1E cells were exposed to the indicated d -glucose (D-Glc) and l -glucose (L-Glc) concentrations for 48 h; during the last 16 h, the cells were incubated with serum-free culture medium with the same additions. The media (10 mL) were then collected, extracted, and analyzed by HPLC. Data are given as nanogram 4-HNE per milligram cellular protein. Representative HPLC tracings are depicted ( inset ) and the arrows point to 4-HNE peaks. Results are mean ± SEM, n = 3–4. * P

Techniques Used: Gas Chromatography, Incubation, High Performance Liquid Chromatography

A model for the dual function of 4-HNE in β-cells. Exposure to high glucose markedly enhances ROS production as well as the activation of cPLA 2 in β-cells by inducing Ser 505 and Ser 515 phosphorylations and the subsequent release of arachidonic acid (ARA) and linoleic acid (LNA) from phospholipids. ROS-mediated peroxidation of these PUFAs results in the generation of 4-HNE. This molecule affects β-cell function in two major ways; when present at nontoxic concentrations, it amplifies insulin secretion in a PPAR-δ–RXR–dependent manner. RXR is activated by cis -retinoic acid (cRA). However, chronic hyperglycemia may lead to an excessive generation of 4-HNE with the accumulation of 4-HNE adducts, causing β-cell dysfunction, characteristic of the advanced stages of type 2 diabetes. (A high-quality color representation of this figure is available in the online issue.)
Figure Legend Snippet: A model for the dual function of 4-HNE in β-cells. Exposure to high glucose markedly enhances ROS production as well as the activation of cPLA 2 in β-cells by inducing Ser 505 and Ser 515 phosphorylations and the subsequent release of arachidonic acid (ARA) and linoleic acid (LNA) from phospholipids. ROS-mediated peroxidation of these PUFAs results in the generation of 4-HNE. This molecule affects β-cell function in two major ways; when present at nontoxic concentrations, it amplifies insulin secretion in a PPAR-δ–RXR–dependent manner. RXR is activated by cis -retinoic acid (cRA). However, chronic hyperglycemia may lead to an excessive generation of 4-HNE with the accumulation of 4-HNE adducts, causing β-cell dysfunction, characteristic of the advanced stages of type 2 diabetes. (A high-quality color representation of this figure is available in the online issue.)

Techniques Used: Activation Assay, Acetylene Reduction Assay, Cell Function Assay

High 4-HNE concentrations compromise β-cell survival. A : Rat islets and INS-1E cells were incubated for 48 or 24 h, respectively, with increasing concentrations of 4-HNE. Islets were pooled from three animals and divided into 10–12 islets per group. After incubation, islet cells were dispersed by mild trypsin digestion. Cell viability was determined at the end of the incubation by the trypan blue exclusion test. Cell viability in the absence of 4-HNE was > 95% in both preparations. Results are mean ± SEM, n = 3. * P
Figure Legend Snippet: High 4-HNE concentrations compromise β-cell survival. A : Rat islets and INS-1E cells were incubated for 48 or 24 h, respectively, with increasing concentrations of 4-HNE. Islets were pooled from three animals and divided into 10–12 islets per group. After incubation, islet cells were dispersed by mild trypsin digestion. Cell viability was determined at the end of the incubation by the trypan blue exclusion test. Cell viability in the absence of 4-HNE was > 95% in both preparations. Results are mean ± SEM, n = 3. * P

Techniques Used: Incubation

High glucose and 4-HNE activate PPAR-δ in INS-1E cells. A : INS-1E cells were transfected with the hPPAR-α, hPPAR-γ1, hPPAR-γ2, or hPPAR-δ expression vectors; the hRXR expression vector; and the 3XPPRE-TK-luciferase reporter plasmid. Renilla luciferase plasmid was transfected for normalizing the luciferase activity data. The transfected cells were treated with 60 μmol/L WY14643, 30 μmol/L troglitazone, 0.1 μmol/L GW501516 (GW), or 1.0 μmol/L 4-HNE for 24 h and luciferase activity was then measured. The 100% values were assigned to the respective untreated control groups. * P
Figure Legend Snippet: High glucose and 4-HNE activate PPAR-δ in INS-1E cells. A : INS-1E cells were transfected with the hPPAR-α, hPPAR-γ1, hPPAR-γ2, or hPPAR-δ expression vectors; the hRXR expression vector; and the 3XPPRE-TK-luciferase reporter plasmid. Renilla luciferase plasmid was transfected for normalizing the luciferase activity data. The transfected cells were treated with 60 μmol/L WY14643, 30 μmol/L troglitazone, 0.1 μmol/L GW501516 (GW), or 1.0 μmol/L 4-HNE for 24 h and luciferase activity was then measured. The 100% values were assigned to the respective untreated control groups. * P

Techniques Used: Transfection, Expressing, Plasmid Preparation, Luciferase, Activity Assay

4-HNE mimics high glucose amplification of insulin secretion. A : Rat islets and INS-1E cells were incubated with RPMI-1640 medium containing the indicated glucose levels in the absence or presence of 1 μmol/L GSK0660 for 48 and 24 h, respectively. GSIS was evaluated by 1-h static incubations at 3.3 mmol/L glucose (white bars), followed by a 1-h incubation at 16.7 mmol/L glucose (black bars). Insulin secretion is presented as percent of insulin content. Glc, glucose. Results are mean ± SEM, n = 3. *P
Figure Legend Snippet: 4-HNE mimics high glucose amplification of insulin secretion. A : Rat islets and INS-1E cells were incubated with RPMI-1640 medium containing the indicated glucose levels in the absence or presence of 1 μmol/L GSK0660 for 48 and 24 h, respectively. GSIS was evaluated by 1-h static incubations at 3.3 mmol/L glucose (white bars), followed by a 1-h incubation at 16.7 mmol/L glucose (black bars). Insulin secretion is presented as percent of insulin content. Glc, glucose. Results are mean ± SEM, n = 3. *P

Techniques Used: Amplification, Incubation, Gas Chromatography

38) Product Images from "Haem-activated promiscuous targeting of artemisinin in Plasmodium falciparum"

Article Title: Haem-activated promiscuous targeting of artemisinin in Plasmodium falciparum

Journal: Nature Communications

doi: 10.1038/ncomms10111

In vitro binding and functional validation of artemisinin targets. ( a ) Artemisinin specifically interacts with OAT, PyrK, LDH, SpdSyn, SAMS and TCTP as the unlabelled artesunate (25 ×) can compete with the AP1 binding. Heat denaturation reduces the AP1 -labelling level of OAT, suggesting that the interaction of artemisinin with OAT is activity based. ( b ) Dose-dependent labelling of OAT with AP1 (4 h treatment). ( c ) Time-dependent labelling of OAT with AP1 . ( d ) The interaction of artemisinin with OAT may involve thiol and amine groups as IAA (blocking thiol, 30 mM) and NEM (blocking amine, 10 mM) pretreatment (20 min) can reduce binding. ( e , f ) Activated artesunate inhibits the activities of PyrK ( e ) and LDH ( f ) in vitro . Δ, heat denaturation; IAA, iodoacetamide; NEM, N -ethylmaleimide; Conc., concentration. Error bars represent s.d. in three independent replicates in e and f . Full-gel images for panels a – d are shown in Supplementary Fig. 13 .
Figure Legend Snippet: In vitro binding and functional validation of artemisinin targets. ( a ) Artemisinin specifically interacts with OAT, PyrK, LDH, SpdSyn, SAMS and TCTP as the unlabelled artesunate (25 ×) can compete with the AP1 binding. Heat denaturation reduces the AP1 -labelling level of OAT, suggesting that the interaction of artemisinin with OAT is activity based. ( b ) Dose-dependent labelling of OAT with AP1 (4 h treatment). ( c ) Time-dependent labelling of OAT with AP1 . ( d ) The interaction of artemisinin with OAT may involve thiol and amine groups as IAA (blocking thiol, 30 mM) and NEM (blocking amine, 10 mM) pretreatment (20 min) can reduce binding. ( e , f ) Activated artesunate inhibits the activities of PyrK ( e ) and LDH ( f ) in vitro . Δ, heat denaturation; IAA, iodoacetamide; NEM, N -ethylmaleimide; Conc., concentration. Error bars represent s.d. in three independent replicates in e and f . Full-gel images for panels a – d are shown in Supplementary Fig. 13 .

Techniques Used: In Vitro, Binding Assay, Functional Assay, Activity Assay, Blocking Assay, Concentration Assay

39) Product Images from "Pharmacokinetic Interaction of Green Rooibos Extract With Atorvastatin and Metformin in Rats"

Article Title: Pharmacokinetic Interaction of Green Rooibos Extract With Atorvastatin and Metformin in Rats

Journal: Frontiers in Pharmacology

doi: 10.3389/fphar.2019.01243

Semi-log plot of the plasma concentration versus time of (A) atorvastatin and (B) metformin in the absence and presence of GRT in Wistar rats (n = 24 per group). The black line indicates atorvastatin or metformin, and gray lines indicate atorvastatin with GRT and metformin with GRT. Ator, atorvastatin; Met, metformin; GRT, green rooibos extract.
Figure Legend Snippet: Semi-log plot of the plasma concentration versus time of (A) atorvastatin and (B) metformin in the absence and presence of GRT in Wistar rats (n = 24 per group). The black line indicates atorvastatin or metformin, and gray lines indicate atorvastatin with GRT and metformin with GRT. Ator, atorvastatin; Met, metformin; GRT, green rooibos extract.

Techniques Used: Concentration Assay

40) Product Images from "Endothelium-Dependent Vasorelaxant Effect of Butanolic Fraction from Caryocar brasiliense Camb. Leaves in Rat Thoracic Aorta"

Article Title: Endothelium-Dependent Vasorelaxant Effect of Butanolic Fraction from Caryocar brasiliense Camb. Leaves in Rat Thoracic Aorta

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2012/934142

Relaxation evoked by BF before (control curve) and after exposure to pyrilamine (a H 1 receptor antagonist; 10 μ M) or atropine (a muscarinic antagonist receptor; 10 μ M) in rat aortic rings. The results are expressed as mean ± SEM of 5–7 experiments.
Figure Legend Snippet: Relaxation evoked by BF before (control curve) and after exposure to pyrilamine (a H 1 receptor antagonist; 10 μ M) or atropine (a muscarinic antagonist receptor; 10 μ M) in rat aortic rings. The results are expressed as mean ± SEM of 5–7 experiments.

Techniques Used:

41) Product Images from "Liquid chromatography electrospray ionization tandem mass spectrometry analysis method for simultaneous detection of trichloroacetic acid, dichloroacetic acid, S-(1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-L-cysteine"

Article Title: Liquid chromatography electrospray ionization tandem mass spectrometry analysis method for simultaneous detection of trichloroacetic acid, dichloroacetic acid, S-(1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-L-cysteine

Journal: Toxicology

doi: 10.1016/j.tox.2009.06.013

MS/MS product ion spectra of [M-H] - ions of chemical standards. (A) dichloroacetic acid, (B) difluoroacetic acid, (C) trichloroacetic acid, and (D) trifluoroacetic acid.
Figure Legend Snippet: MS/MS product ion spectra of [M-H] - ions of chemical standards. (A) dichloroacetic acid, (B) difluoroacetic acid, (C) trichloroacetic acid, and (D) trifluoroacetic acid.

Techniques Used: Mass Spectrometry

42) Product Images from "Lactobacillus plantarum IS-10506 probiotic administration increases amlodipine absorption in a rabbit model"

Article Title: Lactobacillus plantarum IS-10506 probiotic administration increases amlodipine absorption in a rabbit model

Journal: The Journal of International Medical Research

doi: 10.1177/0300060518788994

Chromatogram of amlodipine and nortriptyline as an internal standard in rabbit plasma AML: amlodipine; NOR: nortriptyline hydrochloride.
Figure Legend Snippet: Chromatogram of amlodipine and nortriptyline as an internal standard in rabbit plasma AML: amlodipine; NOR: nortriptyline hydrochloride.

Techniques Used:

43) Product Images from "Density Functional Theory Analysis of Deltamethrin and Its Determination in Strawberry by Surface Enhanced Raman Spectroscopy"

Article Title: Density Functional Theory Analysis of Deltamethrin and Its Determination in Strawberry by Surface Enhanced Raman Spectroscopy

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

doi: 10.3390/molecules23061458

( a ) 500–1800 cm −1 SERS spectra of different concentrations of deltamethrin in strawberry; ( b ) the SERS spectra of five concentrations of deltamethrin in strawberry.
Figure Legend Snippet: ( a ) 500–1800 cm −1 SERS spectra of different concentrations of deltamethrin in strawberry; ( b ) the SERS spectra of five concentrations of deltamethrin in strawberry.

Techniques Used:

The RS of deltamethrin: ( a ) the theory calculation by density functional theory; ( b ) RS of deltamethrin solid; ( c ) SERS of deltamethrin solution.
Figure Legend Snippet: The RS of deltamethrin: ( a ) the theory calculation by density functional theory; ( b ) RS of deltamethrin solid; ( c ) SERS of deltamethrin solution.

Techniques Used: Functional Assay

The SERS of deltamethrin solution based on two nanoparticles: (a) AgNPs; (b) AuNPs; the SERS of two nanoparticles; (c) AgNPs; (d) AuNPs; the RS of acetonitrile; (e) acetonitrile.
Figure Legend Snippet: The SERS of deltamethrin solution based on two nanoparticles: (a) AgNPs; (b) AuNPs; the SERS of two nanoparticles; (c) AgNPs; (d) AuNPs; the RS of acetonitrile; (e) acetonitrile.

Techniques Used:

Raman shift deviation between the Raman characteristic peaks of deltamethrin and the Raman characteristic peaks calculated by DFT.
Figure Legend Snippet: Raman shift deviation between the Raman characteristic peaks of deltamethrin and the Raman characteristic peaks calculated by DFT.

Techniques Used:

Simulated molecular structure of deltamethrin by DFT.
Figure Legend Snippet: Simulated molecular structure of deltamethrin by DFT.

Techniques Used:

44) Product Images from "Aldehyde Oxidase Functions as a Superoxide Generating NADH Oxidase: An Important Redox Regulated Pathway of Cellular Oxygen Radical Formation"

Article Title: Aldehyde Oxidase Functions as a Superoxide Generating NADH Oxidase: An Important Redox Regulated Pathway of Cellular Oxygen Radical Formation

Journal: Biochemistry

doi: 10.1021/bi3000879

Cytochrome c assay for the generation of O 2 • − during the oxidation of NADH by AO
Figure Legend Snippet: Cytochrome c assay for the generation of O 2 • − during the oxidation of NADH by AO

Techniques Used:

45) Product Images from "Renal Damaging Effect Elicited by Bicalutamide Therapy Uncovered Multiple Action Mechanisms As Evidenced by the Cell Model"

Article Title: Renal Damaging Effect Elicited by Bicalutamide Therapy Uncovered Multiple Action Mechanisms As Evidenced by the Cell Model

Journal: Scientific Reports

doi: 10.1038/s41598-019-39533-3

Late apoptosis of RMC cell lines affected by treatment of bicalutamide. ( a ) The apoptosis profile affected by treating with bicalutamide respectively at 24 and 48 h when examined with Annexin V and 7-ADD staining. ( b ) The quantified cell population in each quadrant of ( a ) at 24 h. ( c ) The quantified cell population in each quadrant of ( a ) at 48 h. ( d ) The representative protein expression of autophagic marker LC3B-I and LC3B-II after Bic treatment for 48 h. Late apoptosis of RMC cell lines affected by bicalutamide. Experiment was performed in triplicate (n = 3).
Figure Legend Snippet: Late apoptosis of RMC cell lines affected by treatment of bicalutamide. ( a ) The apoptosis profile affected by treating with bicalutamide respectively at 24 and 48 h when examined with Annexin V and 7-ADD staining. ( b ) The quantified cell population in each quadrant of ( a ) at 24 h. ( c ) The quantified cell population in each quadrant of ( a ) at 48 h. ( d ) The representative protein expression of autophagic marker LC3B-I and LC3B-II after Bic treatment for 48 h. Late apoptosis of RMC cell lines affected by bicalutamide. Experiment was performed in triplicate (n = 3).

Techniques Used: Staining, Expressing, Marker

LC/MS/MS analysis of testosterone. ( a ) The standard curve of testosterone. ( b ) The standard curve of bicalutamide. ( c ) The time–dependent change of extracellular concentration of testosterone.
Figure Legend Snippet: LC/MS/MS analysis of testosterone. ( a ) The standard curve of testosterone. ( b ) The standard curve of bicalutamide. ( c ) The time–dependent change of extracellular concentration of testosterone.

Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Concentration Assay

Dose dependent effect of co-treatment with testosterone and bicalutamide on the survival and apoptotic pathways in the RMC cells. ( a ) TNF-α and TNFR. ( b ) Total and phosphorated PI3K, PDGF-R, and Akt. ( c ) Caspase-3 and cleaved caspase-3. Experiment was performed in triplicate and statistically treated (n = 3). The symbol ‘*’ compares within the same group; and ‘#’ compares among groups. * p
Figure Legend Snippet: Dose dependent effect of co-treatment with testosterone and bicalutamide on the survival and apoptotic pathways in the RMC cells. ( a ) TNF-α and TNFR. ( b ) Total and phosphorated PI3K, PDGF-R, and Akt. ( c ) Caspase-3 and cleaved caspase-3. Experiment was performed in triplicate and statistically treated (n = 3). The symbol ‘*’ compares within the same group; and ‘#’ compares among groups. * p

Techniques Used:

Expression of signaling proteins and Annexin V% affected by testosterone and bicalutamide. ( a ) PI3K and Akt proteins. ( b ) The expression of p-Akt and total Akt when treated as indicated. ( c ) Apoptotic effect of SC79 and a combination of SC79 plus MK2206. The dose(s) used were: testosterone (T) 100 nM; bicalutamide (7.5, 15, 30 and 60 μM); SC79 10 μM, and MK2206 1 μM. Triplicate experiments were statistically treated. Data expressed in mean ± S.D. (n = 3).
Figure Legend Snippet: Expression of signaling proteins and Annexin V% affected by testosterone and bicalutamide. ( a ) PI3K and Akt proteins. ( b ) The expression of p-Akt and total Akt when treated as indicated. ( c ) Apoptotic effect of SC79 and a combination of SC79 plus MK2206. The dose(s) used were: testosterone (T) 100 nM; bicalutamide (7.5, 15, 30 and 60 μM); SC79 10 μM, and MK2206 1 μM. Triplicate experiments were statistically treated. Data expressed in mean ± S.D. (n = 3).

Techniques Used: Expressing

Effect of bicalutamide and TGF-β on the collagen synthesis and associated fibrogenic proteins in RMC cells. ( a ) Transwell assay staining of the control, TGB-β-treated, and bicalutamide-treated RMC cells. ( b ) Quantification of the cell numbers on the transwell assay. ( c ) Quantification of sirius red for the collagen synthesis (numeral data was shown as Supplementary Table 1 ); and ( d ) Western blot on the expression of fibrogenic proteins PDGF-β, fibronectin, collagen IV, and SMA. Experiments were performed in triplicate and statistically treated (n = 3). The dose of bicalutamide used was 7.5, 15, 30 and 60 μM, respectively. T means Testosterone 100 nM.
Figure Legend Snippet: Effect of bicalutamide and TGF-β on the collagen synthesis and associated fibrogenic proteins in RMC cells. ( a ) Transwell assay staining of the control, TGB-β-treated, and bicalutamide-treated RMC cells. ( b ) Quantification of the cell numbers on the transwell assay. ( c ) Quantification of sirius red for the collagen synthesis (numeral data was shown as Supplementary Table 1 ); and ( d ) Western blot on the expression of fibrogenic proteins PDGF-β, fibronectin, collagen IV, and SMA. Experiments were performed in triplicate and statistically treated (n = 3). The dose of bicalutamide used was 7.5, 15, 30 and 60 μM, respectively. T means Testosterone 100 nM.

Techniques Used: Transwell Assay, Staining, Western Blot, Expressing

The cell cycle analysis of RMC cell line affected by bicalutamide treatment. ( a ) Histogram for flowcytometric analysis. ( b ) Quantification of cell populations in 24 h. ( c ) Quantification of cell populations in 48 h. Experiment was performed in triplicate
Figure Legend Snippet: The cell cycle analysis of RMC cell line affected by bicalutamide treatment. ( a ) Histogram for flowcytometric analysis. ( b ) Quantification of cell populations in 24 h. ( c ) Quantification of cell populations in 48 h. Experiment was performed in triplicate

Techniques Used: Cell Cycle Assay

The representative expressions and the quantifications of proteins regarding the bicalutamide-induced apoptosis. ( a ) downregulation of nuclear androgen receptor (nAR), upregulation of ( b ) TNF-α, ( c ) cytosolic and nuclear NFκB p50, ( d ) pro- and cleaved- caspase 3, but not via ( e ) Fas. Experiment was performed in triplicate (n = 3). LG: low glucose 5.5 mM; T: Testosterone 100 nM.
Figure Legend Snippet: The representative expressions and the quantifications of proteins regarding the bicalutamide-induced apoptosis. ( a ) downregulation of nuclear androgen receptor (nAR), upregulation of ( b ) TNF-α, ( c ) cytosolic and nuclear NFκB p50, ( d ) pro- and cleaved- caspase 3, but not via ( e ) Fas. Experiment was performed in triplicate (n = 3). LG: low glucose 5.5 mM; T: Testosterone 100 nM.

Techniques Used:

MTT assay of NRK52E- and RMC cells affected by testosterone, bicalutamide, and R1881 in 25 mM hyperglycemic medium. NRK52E cell line (1 × 10 4 cells/well) on 24-well plate was incubated in medium containing 5% bovine serum and 0.4% PBS overnight until adhered. Finally, the medium was replaced fresh with serum free medium. RMC cell line (1 × 10 4 cells/well) on 24-well plate was incubated in medium containing 25 mM glucose, 15% FBS, 0.4% PBS, and 0.8% G418 overnight until adhered. Finally, the medium was replaced fresh with 2% FBS. At final step, the two cells were simultaneously treated as indicated. ( a ) NRK52E + testosterone (1~10 3 nM), ( b ) NRK52E + bicalutamide (3.75~60 μM), ( c ) RMC + testosterone (1~10 3 nM), ( d ) RMC + R1881 (1~10 3 nM), and ( e ) RMC + bicalutamide, and ( f ) RMC + bicalutamide + testosterone for duration as indicated. Experiment was performed in triplicate
Figure Legend Snippet: MTT assay of NRK52E- and RMC cells affected by testosterone, bicalutamide, and R1881 in 25 mM hyperglycemic medium. NRK52E cell line (1 × 10 4 cells/well) on 24-well plate was incubated in medium containing 5% bovine serum and 0.4% PBS overnight until adhered. Finally, the medium was replaced fresh with serum free medium. RMC cell line (1 × 10 4 cells/well) on 24-well plate was incubated in medium containing 25 mM glucose, 15% FBS, 0.4% PBS, and 0.8% G418 overnight until adhered. Finally, the medium was replaced fresh with 2% FBS. At final step, the two cells were simultaneously treated as indicated. ( a ) NRK52E + testosterone (1~10 3 nM), ( b ) NRK52E + bicalutamide (3.75~60 μM), ( c ) RMC + testosterone (1~10 3 nM), ( d ) RMC + R1881 (1~10 3 nM), and ( e ) RMC + bicalutamide, and ( f ) RMC + bicalutamide + testosterone for duration as indicated. Experiment was performed in triplicate

Techniques Used: MTT Assay, Incubation

Effect of bicalutamide on expression of proteins related with the intrinsic and ER stress pathways. ( a ) The intrinsic pathway related proteins Bcl-2 and Bax. ( b ) The ER–associated calpain-1 and caspase-12 proteins. Western blots were performed in triplicate and quantified (n = 3). LG: low glucose 5.5 mM; E: Etoptoside 2 μM.
Figure Legend Snippet: Effect of bicalutamide on expression of proteins related with the intrinsic and ER stress pathways. ( a ) The intrinsic pathway related proteins Bcl-2 and Bax. ( b ) The ER–associated calpain-1 and caspase-12 proteins. Western blots were performed in triplicate and quantified (n = 3). LG: low glucose 5.5 mM; E: Etoptoside 2 μM.

Techniques Used: Expressing, Western Blot

46) Product Images from "A pilot study of the modulation of sirtuins on arylamine N-acetyltransferase 1 and 2 enzymatic activity"

Article Title: A pilot study of the modulation of sirtuins on arylamine N-acetyltransferase 1 and 2 enzymatic activity

Journal: Acta Pharmaceutica Sinica. B

doi: 10.1016/j.apsb.2017.11.008

Dose-response and kinetic curves for the detection of acetyl- p -aminobenzoic acid (Ac-PABA) and acetyl-isoniazid (Ac-INH) in peripheral blood mononuclear cells (PBMC) and HeLa cells. HeLa cells (A), (B) and PBMC (C), (D) were incubated in the presence of different concentrations of PABA (A), (C) or INH (B), (D). Both metabolites were quantified by HPLC as described in Section materials and methods. Each point represents the mean of duplicate assays performed in four cultures. M, mol/L.
Figure Legend Snippet: Dose-response and kinetic curves for the detection of acetyl- p -aminobenzoic acid (Ac-PABA) and acetyl-isoniazid (Ac-INH) in peripheral blood mononuclear cells (PBMC) and HeLa cells. HeLa cells (A), (B) and PBMC (C), (D) were incubated in the presence of different concentrations of PABA (A), (C) or INH (B), (D). Both metabolites were quantified by HPLC as described in Section materials and methods. Each point represents the mean of duplicate assays performed in four cultures. M, mol/L.

Techniques Used: Incubation, High Performance Liquid Chromatography

Effect of an inhibitor of sirtuins on NAT1 and NAT2 activity. Peripheral blood mononuclear cells (C) from healthy subjects were cultured with different concentrations of a sirtuin inhibitor (nicotinamide, NAM: 0, 30, 100 µmol/L, 5 and 20 mmol/L) for 3 h and, subsequently, the substrate of each enzyme with p -aminobenzoic acid (PABA) or isoniazid (INH) were added. NAT1 (A) and NAT2 (B) activity expressed in nmol of metabolite/min/mg of protein was determined by HPLC. PABA or INH concentrations = 10, 30 and 100 µmol/L. * P
Figure Legend Snippet: Effect of an inhibitor of sirtuins on NAT1 and NAT2 activity. Peripheral blood mononuclear cells (C) from healthy subjects were cultured with different concentrations of a sirtuin inhibitor (nicotinamide, NAM: 0, 30, 100 µmol/L, 5 and 20 mmol/L) for 3 h and, subsequently, the substrate of each enzyme with p -aminobenzoic acid (PABA) or isoniazid (INH) were added. NAT1 (A) and NAT2 (B) activity expressed in nmol of metabolite/min/mg of protein was determined by HPLC. PABA or INH concentrations = 10, 30 and 100 µmol/L. * P

Techniques Used: Activity Assay, Cell Culture, High Performance Liquid Chromatography

NAT1 and NAT2 enzymatic activity in mononuclear cell cultures. Peripheral blood mononuclear cells (PBMC) were cultured with 10, 30 and 100 µmol/L of PABA (A) or INH (B) for 24 h. (C) Comparison of enzymatic activity between NAT1 (■) and NAT2 (•). Levels of Ac-PABA and Ac-INH from each culture were evaluated by HPLC as described in Section materials and methods. Each point represents the mean of duplicate assays of cultured cells. (D) Correlation between the percentage of NAT2 positive cells and NAT2 activity measured as acetyl-isoniazid (Ac-INH). A Pearson correlation analysis was performed for each pair; the trend line, correlation coefficient ( r ), and significance ( P ) are shown for the plot ( * P
Figure Legend Snippet: NAT1 and NAT2 enzymatic activity in mononuclear cell cultures. Peripheral blood mononuclear cells (PBMC) were cultured with 10, 30 and 100 µmol/L of PABA (A) or INH (B) for 24 h. (C) Comparison of enzymatic activity between NAT1 (■) and NAT2 (•). Levels of Ac-PABA and Ac-INH from each culture were evaluated by HPLC as described in Section materials and methods. Each point represents the mean of duplicate assays of cultured cells. (D) Correlation between the percentage of NAT2 positive cells and NAT2 activity measured as acetyl-isoniazid (Ac-INH). A Pearson correlation analysis was performed for each pair; the trend line, correlation coefficient ( r ), and significance ( P ) are shown for the plot ( * P

Techniques Used: Activity Assay, Cell Culture, High Performance Liquid Chromatography

Effect of an agonist of sirtuins on NAT1 and NAT2 activity. Peripheral blood mononuclear cells (PBMC) were cultured with different concentrations of a sirtuin agonist (resveratrol, RSV: 0, 10, 50 and 100 µmol/L) for 3 h and, subsequently, with p -aminobenzoic acid (PABA) or isoniazid (INH) were added. NAT1 (A) and NAT2 (B) activity expressed in nmol of metabolite/min/mg of protein was determined by HPLC. PABA or INH concentrations = 10, 30 and 100 µmol/L. M, mol/L.
Figure Legend Snippet: Effect of an agonist of sirtuins on NAT1 and NAT2 activity. Peripheral blood mononuclear cells (PBMC) were cultured with different concentrations of a sirtuin agonist (resveratrol, RSV: 0, 10, 50 and 100 µmol/L) for 3 h and, subsequently, with p -aminobenzoic acid (PABA) or isoniazid (INH) were added. NAT1 (A) and NAT2 (B) activity expressed in nmol of metabolite/min/mg of protein was determined by HPLC. PABA or INH concentrations = 10, 30 and 100 µmol/L. M, mol/L.

Techniques Used: Activity Assay, Cell Culture, High Performance Liquid Chromatography

47) Product Images from "New Kunitz-Type HCRG Polypeptides from the Sea Anemone Heteractis crispa"

Article Title: New Kunitz-Type HCRG Polypeptides from the Sea Anemone Heteractis crispa

Journal: Marine Drugs

doi: 10.3390/md13106038

Elution profiles of H. crispa polypeptides at various stages of chromatographic purification. ( A ) Gel filtration chromatography of polypeptides contained in 80% acetone powder on column with Akrilex P-4; ( B ) Subsequent cation-exchange chromatography of active fraction polypeptides ( Figure 1 A, peak 3) on column with cellulose CM-32; ( C ) RP-HPLC performed on Nucleosil C 18 column of polypeptides ( Figure 1 B, peak 4) desalted on an Akrilex P-4 column. Fraction with hemolytic and trypsin inhibitory activities are accentuated by solid and dotted lines, respectively. MALDI-TOF/MS spectrums and molecular masses of HCRG1 ( D ) and HCRG2 ( E ) after RP-HPLC are shown in the inset. Chromatography conditions are described in the Experimental Section (Methods).
Figure Legend Snippet: Elution profiles of H. crispa polypeptides at various stages of chromatographic purification. ( A ) Gel filtration chromatography of polypeptides contained in 80% acetone powder on column with Akrilex P-4; ( B ) Subsequent cation-exchange chromatography of active fraction polypeptides ( Figure 1 A, peak 3) on column with cellulose CM-32; ( C ) RP-HPLC performed on Nucleosil C 18 column of polypeptides ( Figure 1 B, peak 4) desalted on an Akrilex P-4 column. Fraction with hemolytic and trypsin inhibitory activities are accentuated by solid and dotted lines, respectively. MALDI-TOF/MS spectrums and molecular masses of HCRG1 ( D ) and HCRG2 ( E ) after RP-HPLC are shown in the inset. Chromatography conditions are described in the Experimental Section (Methods).

Techniques Used: Purification, Filtration, Chromatography, High Performance Liquid Chromatography, Mass Spectrometry

48) Product Images from "Smilagenin Protects Dopaminergic Neurons in Chronic MPTP/Probenecid—Lesioned Parkinson’s Disease Models"

Article Title: Smilagenin Protects Dopaminergic Neurons in Chronic MPTP/Probenecid—Lesioned Parkinson’s Disease Models

Journal: Frontiers in Cellular Neuroscience

doi: 10.3389/fncel.2019.00018

Effect of SMI on striatal dopamine D1 and D2 receptor density and protein level in chronic MPTP/probenecid-lesioned mice. (A) D1 receptor density, assessed by 3 H-SCH23390 ( n = 9). Panel (B) D2 receptor density, assessed by 3 H-spiperone ( n = 9). Panel (C) shows representative western blot bands of D1 and D2 receptor. Panels (D,E) show relative protein expression levels were quantified by densitometry analysis using Image J software on D1 and D2 receptor bands ( n = 4). Data are expressed as mean ± SEM. * P
Figure Legend Snippet: Effect of SMI on striatal dopamine D1 and D2 receptor density and protein level in chronic MPTP/probenecid-lesioned mice. (A) D1 receptor density, assessed by 3 H-SCH23390 ( n = 9). Panel (B) D2 receptor density, assessed by 3 H-spiperone ( n = 9). Panel (C) shows representative western blot bands of D1 and D2 receptor. Panels (D,E) show relative protein expression levels were quantified by densitometry analysis using Image J software on D1 and D2 receptor bands ( n = 4). Data are expressed as mean ± SEM. * P

Techniques Used: Mouse Assay, Western Blot, Expressing, Software

49) Product Images from "Cardioprotective effects of iron chelator HAPI and ROS-activated boronate prochelator BHAPI against catecholamine-induced oxidative cellular injury"

Article Title: Cardioprotective effects of iron chelator HAPI and ROS-activated boronate prochelator BHAPI against catecholamine-induced oxidative cellular injury

Journal: Toxicology

doi: 10.1016/j.tox.2016.10.004

Intracellular Fe-chelating efficiency of Fe chelator HAPI and prochelator BHAPI Effects of 24h-preoxidized CA isoprenaline (oxISO) or epinephrine (oxEPI) on prochelator BHAPI inside H9c2 cells were determined during 10 min with Calcein-AM assay. (A, C) Time course of BHAPI activation to effective chelator with (A) oxISO or (C) oxEPI. (B, D) Efficiency of prochelator activation to effective chelator with (B) oxISO or (D) oxEPI at time 10 min. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control, # vs. corresponding Fe chelator.
Figure Legend Snippet: Intracellular Fe-chelating efficiency of Fe chelator HAPI and prochelator BHAPI Effects of 24h-preoxidized CA isoprenaline (oxISO) or epinephrine (oxEPI) on prochelator BHAPI inside H9c2 cells were determined during 10 min with Calcein-AM assay. (A, C) Time course of BHAPI activation to effective chelator with (A) oxISO or (C) oxEPI. (B, D) Efficiency of prochelator activation to effective chelator with (B) oxISO or (D) oxEPI at time 10 min. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control, # vs. corresponding Fe chelator.

Techniques Used: Calcein AM Assay, Activation Assay

Comparison of cytotoxic effects of Fe chelator HAPI and prochelator BHAPI towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. H9c2 cells were incubated with increasing concentrations of tested compounds for 24h (A, B) or 72h (C, D) Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group.
Figure Legend Snippet: Comparison of cytotoxic effects of Fe chelator HAPI and prochelator BHAPI towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. H9c2 cells were incubated with increasing concentrations of tested compounds for 24h (A, B) or 72h (C, D) Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group.

Techniques Used: Incubation

Cellular morphology and nuclear epifluorescence staining with Hoechst 33342 and PI H9c2 cardiomyoblasts were incubated for 24 h with (A) control medium, (B) 24h-preoxidized EPI (oxEPI) alone, or in combination with studied compound (C) HAPI or (D) BHAPI, when the compound was added to oxEPI immediately before cellular experiment. Scale bars represent 100 μm.
Figure Legend Snippet: Cellular morphology and nuclear epifluorescence staining with Hoechst 33342 and PI H9c2 cardiomyoblasts were incubated for 24 h with (A) control medium, (B) 24h-preoxidized EPI (oxEPI) alone, or in combination with studied compound (C) HAPI or (D) BHAPI, when the compound was added to oxEPI immediately before cellular experiment. Scale bars represent 100 μm.

Techniques Used: Staining, Incubation

Protective effects of HAPI and BHAPI against oxEPI-induced toxicities towards isolated rat neonatal ventricularcardiomyocytes (NVCM) NVCM were incubated for 24 h with studied compounds or with their combination with 24h-preoxidized EPI (oxEPI; 700 μM). (A) Using epifluorescence microscopy, mitochondrial depolarization was assessed after loading with the JC-1 probe (red emission reflects mitochondrial inner membrane potential-dependent accumulation of probe dimers in actively respiring mitochondria, green fluorescence indicates monomers of the probe released into the cytoplasm after mitochondrial depolarization, lack of fluorescence reflects probe release from necrotic or late-stage apoptotic cells). Scale bars represent 50 μm. (B, C) Inherent toxicities of HAPI and BHAPI determined after 24 h incubation with NVCM by measurement of lactate dehydrogenase release. (D, E) Protective effects of HAPI and BHAPI against oxEPI-induced damage on NVCM assessed by measurement of lactate dehydrogenase release after 24 h incubation. Data are presented as means ± S.D.; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. oxEPI group.
Figure Legend Snippet: Protective effects of HAPI and BHAPI against oxEPI-induced toxicities towards isolated rat neonatal ventricularcardiomyocytes (NVCM) NVCM were incubated for 24 h with studied compounds or with their combination with 24h-preoxidized EPI (oxEPI; 700 μM). (A) Using epifluorescence microscopy, mitochondrial depolarization was assessed after loading with the JC-1 probe (red emission reflects mitochondrial inner membrane potential-dependent accumulation of probe dimers in actively respiring mitochondria, green fluorescence indicates monomers of the probe released into the cytoplasm after mitochondrial depolarization, lack of fluorescence reflects probe release from necrotic or late-stage apoptotic cells). Scale bars represent 50 μm. (B, C) Inherent toxicities of HAPI and BHAPI determined after 24 h incubation with NVCM by measurement of lactate dehydrogenase release. (D, E) Protective effects of HAPI and BHAPI against oxEPI-induced damage on NVCM assessed by measurement of lactate dehydrogenase release after 24 h incubation. Data are presented as means ± S.D.; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. oxEPI group.

Techniques Used: Isolation, Incubation, Epifluorescence Microscopy, Fluorescence

Process of BHAPI activation to HAPI HPLC analyses of spontaneous degradation of BHAPI and HAPI and of effects of freshly-prepared or 24h-preoxidized EPI. BHAPI or HAPI were incubated for 24 h: (A, D, G) alone, (B, E, H) with EPI or (C, F, I) with oxEPI in (A, B, C) buffered solution (pH 7.4), (D, E, F) serum-free cell-culture medium or (G, H, I) serum-free cell-culture medium with H9c2 cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. t 0 .
Figure Legend Snippet: Process of BHAPI activation to HAPI HPLC analyses of spontaneous degradation of BHAPI and HAPI and of effects of freshly-prepared or 24h-preoxidized EPI. BHAPI or HAPI were incubated for 24 h: (A, D, G) alone, (B, E, H) with EPI or (C, F, I) with oxEPI in (A, B, C) buffered solution (pH 7.4), (D, E, F) serum-free cell-culture medium or (G, H, I) serum-free cell-culture medium with H9c2 cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. t 0 .

Techniques Used: Activation Assay, High Performance Liquid Chromatography, Incubation, Cell Culture

Comparison of protective effect of HAPI and BHAPI against ISO- and EPI-induced toxicities towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. (A) HAPI or (B) BHAPI were added to freshly-prepared CA in serum-free cell-culture medium before the start of 24h cellular experiments; (C) HAPI or (D) BHAPI were added immediately before cellular experiments to CA preoxidized for 24 h in serum-free cell-culture medium; (E) HAPI or (F) BHAPI were preincubated for 24 h together with CA in serum-free cell-culture medium and then added to cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.
Figure Legend Snippet: Comparison of protective effect of HAPI and BHAPI against ISO- and EPI-induced toxicities towards H9c2 cardiomyoblast cell line Cellular viabilities were determined by neutral red uptake assay and expressed as a percentage of the untreated control group. (A) HAPI or (B) BHAPI were added to freshly-prepared CA in serum-free cell-culture medium before the start of 24h cellular experiments; (C) HAPI or (D) BHAPI were added immediately before cellular experiments to CA preoxidized for 24 h in serum-free cell-culture medium; (E) HAPI or (F) BHAPI were preincubated for 24 h together with CA in serum-free cell-culture medium and then added to cells. Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.

Techniques Used: Cell Culture

Comparison of effects of HAPI and BHAPI on cellular oxidative stress induced by 24h-preoxidized CA (oxCA – EPI and ISO) Intracellular ROS formation was determined by H 2 DCF-DA assay following the 30min treatment of H9c2 cardiomyoblasts with combination of oxCA and studied compounds. Induced intracellular fluorescence was expressed as a percentage of the group treated with oxCA alone. Cells were incubated with various concentrations of (A) HAPI or (B) BHAPI added immediately before cellular experiments to medium with oxCA (60 μM). Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.
Figure Legend Snippet: Comparison of effects of HAPI and BHAPI on cellular oxidative stress induced by 24h-preoxidized CA (oxCA – EPI and ISO) Intracellular ROS formation was determined by H 2 DCF-DA assay following the 30min treatment of H9c2 cardiomyoblasts with combination of oxCA and studied compounds. Induced intracellular fluorescence was expressed as a percentage of the group treated with oxCA alone. Cells were incubated with various concentrations of (A) HAPI or (B) BHAPI added immediately before cellular experiments to medium with oxCA (60 μM). Data are presented as means ± S.D. ; n = 4; Statistical significance (ANOVA, p ≤ 0.05): * vs. control group, # vs. corresponding CA group.

Techniques Used: Fluorescence, Incubation

50) Product Images from "Vibrating Sharp-edge Spray Ionization (VSSI) for Voltage-Free Direct Analysis of Samples using Mass Spectrometry"

Article Title: Vibrating Sharp-edge Spray Ionization (VSSI) for Voltage-Free Direct Analysis of Samples using Mass Spectrometry

Journal: Rapid communications in mass spectrometry : RCM

doi: 10.1002/rcm.8232

Mass spectra of a) homocysteine (1 mM); b) Sucrose (1 mM); c) polyananine peptides; d) ubiquitin (1 mM); e) DNA duplex with Hoechst 33342. Sequence: CAAATTTG Solvent: H 2 O:MeCN (1:1) mixture containing 1% HOAc. Capillary Temperature: 250 °C
Figure Legend Snippet: Mass spectra of a) homocysteine (1 mM); b) Sucrose (1 mM); c) polyananine peptides; d) ubiquitin (1 mM); e) DNA duplex with Hoechst 33342. Sequence: CAAATTTG Solvent: H 2 O:MeCN (1:1) mixture containing 1% HOAc. Capillary Temperature: 250 °C

Techniques Used: Sequencing

51) Product Images from "Quantitative determination of doxorubicin in the exosomes of A549/MCF-7 cancer cells and human plasma using ultra performance liquid chromatography-tandem mass spectrometry"

Article Title: Quantitative determination of doxorubicin in the exosomes of A549/MCF-7 cancer cells and human plasma using ultra performance liquid chromatography-tandem mass spectrometry

Journal: Saudi Pharmaceutical Journal : SPJ

doi: 10.1016/j.jsps.2018.05.011

Chemical structure of Doxorubicin [A] and Ketotifen (I.S) [B].
Figure Legend Snippet: Chemical structure of Doxorubicin [A] and Ketotifen (I.S) [B].

Techniques Used:

The product ion spectra of Doxorubicin (A) and Ketotifen (IS) (B).
Figure Legend Snippet: The product ion spectra of Doxorubicin (A) and Ketotifen (IS) (B).

Techniques Used:

52) Product Images from "Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica) Fertilized with Sodium Selenate"

Article Title: Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica) Fertilized with Sodium Selenate

Journal: Molecules

doi: 10.3390/molecules18055221

SeMSC, total glucosinolates and sulforaphane content and myrosinase activity in broccoli fertilized with water (control) or with 100 µM sodium selenate (treated).
Figure Legend Snippet: SeMSC, total glucosinolates and sulforaphane content and myrosinase activity in broccoli fertilized with water (control) or with 100 µM sodium selenate (treated).

Techniques Used: Activity Assay

53) Product Images from "Lipid composition and cell surface hydrophobicity of Candida albicans influence the efficacy of fluconazole–gentamicin treatment. Lipid composition and cell surface hydrophobicity of Candida albicans influence the efficacy of fluconazole–gentamicin treatment"

Article Title: Lipid composition and cell surface hydrophobicity of Candida albicans influence the efficacy of fluconazole–gentamicin treatment. Lipid composition and cell surface hydrophobicity of Candida albicans influence the efficacy of fluconazole–gentamicin treatment

Journal: Yeast (Chichester, England)

doi: 10.1002/yea.3455

( a) Relative CDR1 gene expression in the Candida albicans CAF4‐2 strain compared with the C. albicans CAF2‐1 strain. Statistical analysis was performed by comparing both experiments. (b) Relative CDR1 gene expression in the C. albicans CAF2‐1 and CAF4‐2 strains grown in the following conditions: control without antimicrobial agents, Flc—treated with fluconazole 4 μg/ml, Gent—treated with gentamicin 256 μg/ml, Flc + Gent—simultaneously treated with fluconazole 4 μg/ml and gentamicin 256 μg/ml. Statistical analysis was performed by comparing the CDR1 expression level of treated with untreated strains, separately. Gene expression levels are reported as mean ± SD of 2 −ΔΔCT values ( n = 6), normalized to 1 for CAF2‐1 in (a) or separately to CAF2‐1 and CAF4‐2 in (b). (c) Cdr1p‐dependent rhodamine 6G (R6G) efflux in C. albicans CAF2‐1 and CAF4‐2 shown as the normalized (normalized to = 1 for CAF2‐1 strain) fluorescence intensity of extracellular R6G (mean ± SD , n = 6). (d) R6G efflux in the C. albicans CAF2‐1 and CAF4‐2 strains grown in the following conditions: control without antimicrobial agents, Flc—treated with fluconazole 4 μg/ml, Gent—treated with gentamicin 256 μg/ml, Flc + Gent—simultaneously treated with fluconazole 4 μg/ml and gentamicin 256 μg/ml (normalized to = 1 separately for untreated CAF2‐1 and CAF4‐2 strains; mean ± SD , n = 6). (e) Fluorescence micrographs of the subcellular localization of the Cdr1‐GFP protein in the C. albicans strains, KS052 (CAF2‐1 CDR1‐GFP) and KS068 (CAF4‐2 CDR1‐GFP) in the following conditions: control without antimicrobial agents, treated with fluconazole 4 μg/ml, treated with gentamicin 256 μg/ml or simultaneously treated with fluconazole 4 μg/ml and gentamicin 256 μg/ml. Scale bar = 5 μm. Statistical significance in all cases is presented as follows: * p
Figure Legend Snippet: ( a) Relative CDR1 gene expression in the Candida albicans CAF4‐2 strain compared with the C. albicans CAF2‐1 strain. Statistical analysis was performed by comparing both experiments. (b) Relative CDR1 gene expression in the C. albicans CAF2‐1 and CAF4‐2 strains grown in the following conditions: control without antimicrobial agents, Flc—treated with fluconazole 4 μg/ml, Gent—treated with gentamicin 256 μg/ml, Flc + Gent—simultaneously treated with fluconazole 4 μg/ml and gentamicin 256 μg/ml. Statistical analysis was performed by comparing the CDR1 expression level of treated with untreated strains, separately. Gene expression levels are reported as mean ± SD of 2 −ΔΔCT values ( n = 6), normalized to 1 for CAF2‐1 in (a) or separately to CAF2‐1 and CAF4‐2 in (b). (c) Cdr1p‐dependent rhodamine 6G (R6G) efflux in C. albicans CAF2‐1 and CAF4‐2 shown as the normalized (normalized to = 1 for CAF2‐1 strain) fluorescence intensity of extracellular R6G (mean ± SD , n = 6). (d) R6G efflux in the C. albicans CAF2‐1 and CAF4‐2 strains grown in the following conditions: control without antimicrobial agents, Flc—treated with fluconazole 4 μg/ml, Gent—treated with gentamicin 256 μg/ml, Flc + Gent—simultaneously treated with fluconazole 4 μg/ml and gentamicin 256 μg/ml (normalized to = 1 separately for untreated CAF2‐1 and CAF4‐2 strains; mean ± SD , n = 6). (e) Fluorescence micrographs of the subcellular localization of the Cdr1‐GFP protein in the C. albicans strains, KS052 (CAF2‐1 CDR1‐GFP) and KS068 (CAF4‐2 CDR1‐GFP) in the following conditions: control without antimicrobial agents, treated with fluconazole 4 μg/ml, treated with gentamicin 256 μg/ml or simultaneously treated with fluconazole 4 μg/ml and gentamicin 256 μg/ml. Scale bar = 5 μm. Statistical significance in all cases is presented as follows: * p

Techniques Used: Expressing, Fluorescence

54) Product Images from "Identification and characterization of phenolics and terpenoids from ethanolic extracts of Phyllanthus species by HPLC-ESI-QTOF-MS/MS"

Article Title: Identification and characterization of phenolics and terpenoids from ethanolic extracts of Phyllanthus species by HPLC-ESI-QTOF-MS/MS

Journal: Journal of Pharmaceutical Analysis

doi: 10.1016/j.jpha.2017.01.005

MS/MS spectra of standards (A) rutin (16), (B) quercetin-3,4-di- O -glucoside (26) and (C) kaempferol-3- O -rutinoside (28) at collision energy 35, 20 and 30 eV, respectively.
Figure Legend Snippet: MS/MS spectra of standards (A) rutin (16), (B) quercetin-3,4-di- O -glucoside (26) and (C) kaempferol-3- O -rutinoside (28) at collision energy 35, 20 and 30 eV, respectively.

Techniques Used: Mass Spectrometry

55) Product Images from "Inhibitory Effects of Aschantin on Cytochrome P450 and Uridine 5′-diphospho-glucuronosyltransferase Enzyme Activities in Human Liver Microsomes"

Article Title: Inhibitory Effects of Aschantin on Cytochrome P450 and Uridine 5′-diphospho-glucuronosyltransferase Enzyme Activities in Human Liver Microsomes

Journal: Molecules

doi: 10.3390/molecules21050554

Inhibitory effects of aschantin on UGT1A1-catalyzed SN-38 glucuronidation, UGT1A3-catalyzed chenodeoxycholic acid 24-acyl-glucuronidation, UGT1A4-catalyzed trifluoperazine N -glucuronidation, UGT1A6-catalyzed N -acetylserotonin glucuronidation, UGT1A9-catalyzed mycophenolic acid glucuronidation, and UGT2B7-catalyzed naloxone 3-β- d -glucuronidationby pooled human liver microsomes. The data are means ± SDs ( n = 3).
Figure Legend Snippet: Inhibitory effects of aschantin on UGT1A1-catalyzed SN-38 glucuronidation, UGT1A3-catalyzed chenodeoxycholic acid 24-acyl-glucuronidation, UGT1A4-catalyzed trifluoperazine N -glucuronidation, UGT1A6-catalyzed N -acetylserotonin glucuronidation, UGT1A9-catalyzed mycophenolic acid glucuronidation, and UGT2B7-catalyzed naloxone 3-β- d -glucuronidationby pooled human liver microsomes. The data are means ± SDs ( n = 3).

Techniques Used:

56) Product Images from "FRET-trackable biodegradable HPMA copolymer-epirubicin conjugates for ovarian carcinoma therapy"

Article Title: FRET-trackable biodegradable HPMA copolymer-epirubicin conjugates for ovarian carcinoma therapy

Journal: Journal of controlled release : official journal of the Controlled Release Society

doi: 10.1016/j.jconrel.2015.09.045

Visualization of payload Cy3 release from conjugate P-Cy3-Cy5 in cathepsin B over-expressing A2780 human ovarian cancer cells by FRET. The cells were first incubated with P-Cy3-Cy5 at 37°C for 4 h and then were washed. Half of the cells were fixed
Figure Legend Snippet: Visualization of payload Cy3 release from conjugate P-Cy3-Cy5 in cathepsin B over-expressing A2780 human ovarian cancer cells by FRET. The cells were first incubated with P-Cy3-Cy5 at 37°C for 4 h and then were washed. Half of the cells were fixed

Techniques Used: Expressing, Incubation

(A) Fluorescence spectra of FRET conjugate P-EPI-Cy5 compared with P-EPI, P-Cy5 and their mixture P-Cy5+P-EPI. (B) The change in FRET ratio of P-EPI-Cy5 conjugate revealed effective EPI release in A2780 ovarian cancer cells. The cathepsin B over-expressing
Figure Legend Snippet: (A) Fluorescence spectra of FRET conjugate P-EPI-Cy5 compared with P-EPI, P-Cy5 and their mixture P-Cy5+P-EPI. (B) The change in FRET ratio of P-EPI-Cy5 conjugate revealed effective EPI release in A2780 ovarian cancer cells. The cathepsin B over-expressing

Techniques Used: Fluorescence, Expressing

57) Product Images from "(S)-4-Trimethylsilyl-3-butyn-2-ol as an auxiliary for stereocontrolled synthesis of salinosporamide analogs with modifications at positions C2 and C5"

Article Title: (S)-4-Trimethylsilyl-3-butyn-2-ol as an auxiliary for stereocontrolled synthesis of salinosporamide analogs with modifications at positions C2 and C5

Journal: Bioorganic & medicinal chemistry letters

doi: 10.1016/j.bmcl.2013.09.066

Synthesis of intermediates to 5-cyclopropyl and 5-benzyl analogs of salinosporamide A (compounds 17 and 18 , respectively, )
Figure Legend Snippet: Synthesis of intermediates to 5-cyclopropyl and 5-benzyl analogs of salinosporamide A (compounds 17 and 18 , respectively, )

Techniques Used:

Inhibition of the 20S proteasome by salinosporamide A ( 1 )
Figure Legend Snippet: Inhibition of the 20S proteasome by salinosporamide A ( 1 )

Techniques Used: Inhibition

 numbered to correspond with salinosporamide A.
Figure Legend Snippet: numbered to correspond with salinosporamide A.

Techniques Used:

58) Product Images from "Administration of Asian Herb Bennet (Geum japonicum) Extract Reverses Depressive-Like Behaviors in Mouse Model of Depression Induced by Corticosterone"

Article Title: Administration of Asian Herb Bennet (Geum japonicum) Extract Reverses Depressive-Like Behaviors in Mouse Model of Depression Induced by Corticosterone

Journal: Nutrients

doi: 10.3390/nu11122841

High-performance liquid chromatography (HPLC) chromatogram for standardization of G. japonicum extract. ( A ) HPLC chromatograms for 3, 4, 5-trihydroxybenzaldehyde and ( B ) G. japonicum extract. X-axis, retention time (min); Y-axis, absorbance unit (AU).
Figure Legend Snippet: High-performance liquid chromatography (HPLC) chromatogram for standardization of G. japonicum extract. ( A ) HPLC chromatograms for 3, 4, 5-trihydroxybenzaldehyde and ( B ) G. japonicum extract. X-axis, retention time (min); Y-axis, absorbance unit (AU).

Techniques Used: High Performance Liquid Chromatography

59) Product Images from "An Integrated Multi-Omic Approach to Assess Radiation Injury on the Host-Microbiome Axis"

Article Title: An Integrated Multi-Omic Approach to Assess Radiation Injury on the Host-Microbiome Axis

Journal: Radiation research

doi: 10.1667/RR14306.1

Panel A: Seven fecal metabolite markers known to be of bacterial origin show statistically significant changes in their abundances after irradiation. Glyceric acid, homogentisic acid, glutaconic acid and pipecolic acid show decreasing abundances after 5 and 12 Gy at day 3. This decrease is dose specific as the mice exposed to 12 Gy show greater decrease in the fecal abundance of these metabolites than those exposed to 5 Gy. Hippuric acid, taurin and urobilinogen show increase in their fecal abundances postirradiation in a dose specific pattern. Because these metabolites are products of the gut microbiota, significant changes in their abundances imply changes in the microbial metabolism and a shift toward gut dysbiosis. Panel B: Multidimensional scaling plot showing the separation of fecal metabolomic profile after 5 Gy irradiation throughout the 30 days. The fecal metabolomic signature of the irradiated mice are well separated from the preirradiation mice (control group). In addition, the metabolomic signatures of day 30 and 14 are closer while that of day 3 is further separated and closer to that of the control group. Panel C: The heatmap of top 50 important variables highlights the time dependence of the metabolomic response to 5 Gy irradiation. The two yellow boxes show individual spectral features with gradual decreasing abundances throughout the 30-day study while the features at the bottom of the heatmap show a rapid drop. Several ions from panel C are shown in panel D. Panel D: these individual microbial markers show time-specific responses to 5 Gy irradiation throughout the 30-day study. Sebacic acid and serotonin showed decreasing levels while 2-ketobutyric acid and hypoxanthine show increasing levels throughout the 30-day study at 5 Gy irradiation.
Figure Legend Snippet: Panel A: Seven fecal metabolite markers known to be of bacterial origin show statistically significant changes in their abundances after irradiation. Glyceric acid, homogentisic acid, glutaconic acid and pipecolic acid show decreasing abundances after 5 and 12 Gy at day 3. This decrease is dose specific as the mice exposed to 12 Gy show greater decrease in the fecal abundance of these metabolites than those exposed to 5 Gy. Hippuric acid, taurin and urobilinogen show increase in their fecal abundances postirradiation in a dose specific pattern. Because these metabolites are products of the gut microbiota, significant changes in their abundances imply changes in the microbial metabolism and a shift toward gut dysbiosis. Panel B: Multidimensional scaling plot showing the separation of fecal metabolomic profile after 5 Gy irradiation throughout the 30 days. The fecal metabolomic signature of the irradiated mice are well separated from the preirradiation mice (control group). In addition, the metabolomic signatures of day 30 and 14 are closer while that of day 3 is further separated and closer to that of the control group. Panel C: The heatmap of top 50 important variables highlights the time dependence of the metabolomic response to 5 Gy irradiation. The two yellow boxes show individual spectral features with gradual decreasing abundances throughout the 30-day study while the features at the bottom of the heatmap show a rapid drop. Several ions from panel C are shown in panel D. Panel D: these individual microbial markers show time-specific responses to 5 Gy irradiation throughout the 30-day study. Sebacic acid and serotonin showed decreasing levels while 2-ketobutyric acid and hypoxanthine show increasing levels throughout the 30-day study at 5 Gy irradiation.

Techniques Used: Irradiation, Mouse Assay

60) Product Images from "CYP2D6-CYP2C9 Protein-Protein Interactions and Isoform-Selective Effects on Substrate Binding and Catalysis"

Article Title: CYP2D6-CYP2C9 Protein-Protein Interactions and Isoform-Selective Effects on Substrate Binding and Catalysis

Journal: Drug Metabolism and Disposition

doi: 10.1124/dmd.109.026500

Effect of CYP2C9 on dextromethorphan metabolism by CYP2D6 at 20 and 40 pmol/incubation of CPR. No CYP2C9 (control) and 1:1 CYP2C9:CYP2D6 ratios were tested.
Figure Legend Snippet: Effect of CYP2C9 on dextromethorphan metabolism by CYP2D6 at 20 and 40 pmol/incubation of CPR. No CYP2C9 (control) and 1:1 CYP2C9:CYP2D6 ratios were tested.

Techniques Used: Incubation

61) Product Images from "Antidiabetic effect of green rooibos (Aspalathus linearis) extract in cultured cells and type 2 diabetic model KK-Ay mice"

Article Title: Antidiabetic effect of green rooibos (Aspalathus linearis) extract in cultured cells and type 2 diabetic model KK-Ay mice

Journal: Cytotechnology

doi: 10.1007/s10616-014-9816-y

Effect of GRE and aspalathin on glucose uptake, AMPK and Akt phosphorylation, and GLUT4 translocation in L6 myotubes. a–c Glucose uptake assay was conducted without or with GRE (0–800 µg/ml) or aspalathin (50 µM)
Figure Legend Snippet: Effect of GRE and aspalathin on glucose uptake, AMPK and Akt phosphorylation, and GLUT4 translocation in L6 myotubes. a–c Glucose uptake assay was conducted without or with GRE (0–800 µg/ml) or aspalathin (50 µM)

Techniques Used: Translocation Assay

62) Product Images from "Phytochemical Profiling of Fruit Powders of Twenty Sorbus L. Cultivars"

Article Title: Phytochemical Profiling of Fruit Powders of Twenty Sorbus L. Cultivars

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

doi: 10.3390/molecules23102593

Anthocyanin compound profiles of different  Sorbus  fruit powder samples. a—cyanidin 3- O -galactoside; b—cyanidin 3- O -glucoside; c—cyanidin 3- O -arabinoside; d—peonidin 3- O -arabinoside; e—Malvidin 3- O -arabinoside. 1—‘Alaja Krupnaja’; 2—‘Burka’; 3—‘Businka’; 4—‘Dodong’; 5—‘Esseziani’; 6—‘Fructo Lutea’; 7—‘Granatnaja’; 8—‘Kirsten Pink’; 9—‘Konzentra’; 10—‘Krasnaja Krupnoplodnaja’; 11—‘Likernaja’; 12—‘Miciurinskaja Desertnaja’; 13—‘Nevezinskaja’; 14—‘Pendula Variegata’; 15—‘Red Tip’; 16—‘Rosina Variegata’; 17—‘Rubinovaja’; 18—‘Sorbinka’; 19—‘Titan’; 20—‘White Swan’.
Figure Legend Snippet: Anthocyanin compound profiles of different Sorbus fruit powder samples. a—cyanidin 3- O -galactoside; b—cyanidin 3- O -glucoside; c—cyanidin 3- O -arabinoside; d—peonidin 3- O -arabinoside; e—Malvidin 3- O -arabinoside. 1—‘Alaja Krupnaja’; 2—‘Burka’; 3—‘Businka’; 4—‘Dodong’; 5—‘Esseziani’; 6—‘Fructo Lutea’; 7—‘Granatnaja’; 8—‘Kirsten Pink’; 9—‘Konzentra’; 10—‘Krasnaja Krupnoplodnaja’; 11—‘Likernaja’; 12—‘Miciurinskaja Desertnaja’; 13—‘Nevezinskaja’; 14—‘Pendula Variegata’; 15—‘Red Tip’; 16—‘Rosina Variegata’; 17—‘Rubinovaja’; 18—‘Sorbinka’; 19—‘Titan’; 20—‘White Swan’.

Techniques Used:

63) Product Images from "Potassium Channel Activation Is Involved in the Cardiovascular Effects Induced by Freeze Dried Syzygium jambolanum (Lam.) DC Fruit Juice"

Article Title: Potassium Channel Activation Is Involved in the Cardiovascular Effects Induced by Freeze Dried Syzygium jambolanum (Lam.) DC Fruit Juice

Journal: BioMed Research International

doi: 10.1155/2018/4827461

Concentration-response curves showing the participation of K + channels in the vasorelaxant effect induced by JSJ. (a) Relaxation induced by JSJ in the endothelium-denuded mesenteric artery rings pre-contracted with Phe (1 μ M) in the absence (○; n=6) or in the simultaneous presence of potassium channel blockers for 4-AP (1 mM), glibenclamide (10 μ M), BaCl 2 (30 μ M) and TEA (1 mM) ( ◇ , n=5), or (b) Iberiotoxin 100 nM (◐, n=6), or (c) BaCl 2 (30 μ M) (◑, n=7), or (d) 4-AP 1 mM (◆), (n = 5), or (e) Glibenclamide 10 μ M ( ∇ , n=7). The Values were expressed as mean ± SEM.
Figure Legend Snippet: Concentration-response curves showing the participation of K + channels in the vasorelaxant effect induced by JSJ. (a) Relaxation induced by JSJ in the endothelium-denuded mesenteric artery rings pre-contracted with Phe (1 μ M) in the absence (○; n=6) or in the simultaneous presence of potassium channel blockers for 4-AP (1 mM), glibenclamide (10 μ M), BaCl 2 (30 μ M) and TEA (1 mM) ( ◇ , n=5), or (b) Iberiotoxin 100 nM (◐, n=6), or (c) BaCl 2 (30 μ M) (◑, n=7), or (d) 4-AP 1 mM (◆), (n = 5), or (e) Glibenclamide 10 μ M ( ∇ , n=7). The Values were expressed as mean ± SEM.

Techniques Used: Concentration Assay

64) Product Images from "Human three-dimensional in vitro model of hepatic zonation to predict zonal hepatotoxicity"

Article Title: Human three-dimensional in vitro model of hepatic zonation to predict zonal hepatotoxicity

Journal: Journal of Biological Engineering

doi: 10.1186/s13036-019-0148-5

Development of the in vitro 3D hepatic zonation platform. a Schematic flow of the zonal hepatotoxicity evaluation system using the 3D hepatic zonal channel model. An agarose hydrogel gel containing HepaRG cells was injected into the polyolefin tube, and 3D HepaRG cells were cultured in the gel matrix for 7 days. For natural diffusion, 0.1% DMSO and 9 μM CHIR were located at the left (Inlet 1) and right side (Inlet 2) of the channel, respectively. The 3D hepatic zonal channel can be sliced into several pieces representing zone-1-like, zone-2-like and zone-3-like sections with the installed guide and then treated with hepatotoxic chemicals. b The relative CHIR diffusion in the 3D hepatic zonal channel was monitored for up to 15 days by UV illumination using the ChemiDoc XRS+ Imaging System. The intensity of CHIR across the 3D hepatic zonal channel was imaged using the MATLAB program. c The relative intensity of CHIR according to the distance from Inlet 1 to Inlet 2 was quantified using the MATLAB program and plotted. d The intensity-concentration curves for each time point were analysed with the curve-fitting method, and the linearity was calculated from the limit (lim) of tangent Ɵ approaching 50% of the distances. A tangent value close to 1 indicates the most linearity. On day 7, the CHIR concentration profile was close to a linear diffusion gradient in the 3D hepatic zonal channel
Figure Legend Snippet: Development of the in vitro 3D hepatic zonation platform. a Schematic flow of the zonal hepatotoxicity evaluation system using the 3D hepatic zonal channel model. An agarose hydrogel gel containing HepaRG cells was injected into the polyolefin tube, and 3D HepaRG cells were cultured in the gel matrix for 7 days. For natural diffusion, 0.1% DMSO and 9 μM CHIR were located at the left (Inlet 1) and right side (Inlet 2) of the channel, respectively. The 3D hepatic zonal channel can be sliced into several pieces representing zone-1-like, zone-2-like and zone-3-like sections with the installed guide and then treated with hepatotoxic chemicals. b The relative CHIR diffusion in the 3D hepatic zonal channel was monitored for up to 15 days by UV illumination using the ChemiDoc XRS+ Imaging System. The intensity of CHIR across the 3D hepatic zonal channel was imaged using the MATLAB program. c The relative intensity of CHIR according to the distance from Inlet 1 to Inlet 2 was quantified using the MATLAB program and plotted. d The intensity-concentration curves for each time point were analysed with the curve-fitting method, and the linearity was calculated from the limit (lim) of tangent Ɵ approaching 50% of the distances. A tangent value close to 1 indicates the most linearity. On day 7, the CHIR concentration profile was close to a linear diffusion gradient in the 3D hepatic zonal channel

Techniques Used: In Vitro, Flow Cytometry, Injection, Cell Culture, Diffusion-based Assay, Imaging, Concentration Assay

65) Product Images from "Hypercrosslinking: New approach to porous polymer monolithic capillary columns with large surface area for the highly efficient separation of small molecules"

Article Title: Hypercrosslinking: New approach to porous polymer monolithic capillary columns with large surface area for the highly efficient separation of small molecules

Journal: Journal of chromatography. A

doi: 10.1016/j.chroma.2010.10.100

Effect of hypercrosslinking of poly(styrene-co-vinylbenzyl chloride-co-divinylbenzene) precursor monolith on the specific surface area. Conditions: Polymerization mixture used for the preparation of precursor monolith: 16% styrene + vinylbenzyl chloride,
Figure Legend Snippet: Effect of hypercrosslinking of poly(styrene-co-vinylbenzyl chloride-co-divinylbenzene) precursor monolith on the specific surface area. Conditions: Polymerization mixture used for the preparation of precursor monolith: 16% styrene + vinylbenzyl chloride,

Techniques Used:

66) Product Images from "A Fast and Validated Reversed-Phase HPLC Method for Simultaneous Determination of Simvastatin, Atorvastatin, Telmisartan and Irbesartan in Bulk Drugs and Tablet Formulations"

Article Title: A Fast and Validated Reversed-Phase HPLC Method for Simultaneous Determination of Simvastatin, Atorvastatin, Telmisartan and Irbesartan in Bulk Drugs and Tablet Formulations

Journal: Scientia Pharmaceutica

doi: 10.3390/scipharm86010001

Chemical structures of ( A ) Atorvastatin (ATV); ( B ) Simvastatin (SMV); ( C ) Telmisartan (TLN) and ( D ) Irbesartan (IRB).
Figure Legend Snippet: Chemical structures of ( A ) Atorvastatin (ATV); ( B ) Simvastatin (SMV); ( C ) Telmisartan (TLN) and ( D ) Irbesartan (IRB).

Techniques Used:

Calibration curve showing excellent linearity of the method: ( A ) Irbesartan; ( B ) Atorvastatin; ( C ) Telmisartan and ( D ) Simvastatin.
Figure Legend Snippet: Calibration curve showing excellent linearity of the method: ( A ) Irbesartan; ( B ) Atorvastatin; ( C ) Telmisartan and ( D ) Simvastatin.

Techniques Used:

Representative chromatograms of individual analytes in tablet dosage forms. ( A ) Irbesartan; ( B ) Atorvastatin; ( C ) Telmisartan; and ( D ) Simvastatin. Conditions : stationary phase, Symmetry C18 column; mobile phase, 10 mM ammonium acetate buffer (pH 4)–acetonitrile (40:60 v / v ); flow rate, 1 mL/min up to 3.5 min then 2 mL/min; detection, UV 220 nm.
Figure Legend Snippet: Representative chromatograms of individual analytes in tablet dosage forms. ( A ) Irbesartan; ( B ) Atorvastatin; ( C ) Telmisartan; and ( D ) Simvastatin. Conditions : stationary phase, Symmetry C18 column; mobile phase, 10 mM ammonium acetate buffer (pH 4)–acetonitrile (40:60 v / v ); flow rate, 1 mL/min up to 3.5 min then 2 mL/min; detection, UV 220 nm.

Techniques Used: Flow Cytometry

Chromatogram showing excellent separation between irbesartan, atorvastatin, telmisartan and simvastatin. Conditions: stationary phase, Symmetry C18 column; mobile phase, 10 mM ammonium acetate buffer (pH 4)–acetonitrile (40:60 v / v ); flow rate, 1 mL/min up to 3.5 min then 2 mL/min; detection, UV 220 nm.
Figure Legend Snippet: Chromatogram showing excellent separation between irbesartan, atorvastatin, telmisartan and simvastatin. Conditions: stationary phase, Symmetry C18 column; mobile phase, 10 mM ammonium acetate buffer (pH 4)–acetonitrile (40:60 v / v ); flow rate, 1 mL/min up to 3.5 min then 2 mL/min; detection, UV 220 nm.

Techniques Used: Flow Cytometry

67) Product Images from "Iridoid Glycosides Fraction Isolated from Veronica ciliata Fisch. Protects against Acetaminophen-Induced Liver Injury in Mice"

Article Title: Iridoid Glycosides Fraction Isolated from Veronica ciliata Fisch. Protects against Acetaminophen-Induced Liver Injury in Mice

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2017/6106572

Antioxidant properties of the iridoid glycosides fraction (IGF) were determined using (a) a DPPH radical scavenging assay; (b) an ABTS radical scavenging assay; (c) a nitrite-scavenging assay; and (d) a reducing power assay. Data are expressed as the mean ± SD ( n = 3). Vc, ascorbic acid; BHT, 2,6-di-tert-butyl-4-methylphenol.
Figure Legend Snippet: Antioxidant properties of the iridoid glycosides fraction (IGF) were determined using (a) a DPPH radical scavenging assay; (b) an ABTS radical scavenging assay; (c) a nitrite-scavenging assay; and (d) a reducing power assay. Data are expressed as the mean ± SD ( n = 3). Vc, ascorbic acid; BHT, 2,6-di-tert-butyl-4-methylphenol.

Techniques Used: DPPH Radical Scavenging Assay

68) Product Images from "An in-vivo pilot study into the effects of FDG-mNP in cancer in mice"

Article Title: An in-vivo pilot study into the effects of FDG-mNP in cancer in mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0202482

Growth profiles of mice injected at 3 weeks of age with 8mk/kg ICG-conjugated FDG-mNPs (A). Representative fluorescent images of kidney (B, D) and brain (C, E) tissue sections from 3 and 6 month old mice injected with ICG-conjugated FDG-mNPs. Tissue intensity (arbitrary units) staining for ICG-conjugated FDG-mNPs normalised to nuclear Hoechst signal in 3 and 6 month old mice (F). n = 1 mouse per treatment group. Images in B-E are at 10x magnification. FDG: fluorodeoxyglucose; ICG: indocyanine; mNP: magnetic nanoparticle.
Figure Legend Snippet: Growth profiles of mice injected at 3 weeks of age with 8mk/kg ICG-conjugated FDG-mNPs (A). Representative fluorescent images of kidney (B, D) and brain (C, E) tissue sections from 3 and 6 month old mice injected with ICG-conjugated FDG-mNPs. Tissue intensity (arbitrary units) staining for ICG-conjugated FDG-mNPs normalised to nuclear Hoechst signal in 3 and 6 month old mice (F). n = 1 mouse per treatment group. Images in B-E are at 10x magnification. FDG: fluorodeoxyglucose; ICG: indocyanine; mNP: magnetic nanoparticle.

Techniques Used: Mouse Assay, Injection, Staining

Representative Haemotoxylin and Eosin stained sections from mice injected with ICG-conjugated FDG-mNPs at a concentration of 8mk/kg. Sections show representative images of brain (A), heart (B), kidney (C), liver (D), lung (E), spleen (F) and muscle (G) in a 3 month old mouse. Sections also show representative images of brain (H), heart (I), kidney (J), liver (K), lung (L), spleen (M) and muscle (N) in a 6 month old mouse. All images are at 10x magnification. FDG: fluorodeoxyglucose; ICG: indocyanine; mNP: magnetic nanoparticle.
Figure Legend Snippet: Representative Haemotoxylin and Eosin stained sections from mice injected with ICG-conjugated FDG-mNPs at a concentration of 8mk/kg. Sections show representative images of brain (A), heart (B), kidney (C), liver (D), lung (E), spleen (F) and muscle (G) in a 3 month old mouse. Sections also show representative images of brain (H), heart (I), kidney (J), liver (K), lung (L), spleen (M) and muscle (N) in a 6 month old mouse. All images are at 10x magnification. FDG: fluorodeoxyglucose; ICG: indocyanine; mNP: magnetic nanoparticle.

Techniques Used: Staining, Mouse Assay, Injection, Concentration Assay

69) Product Images from "Protective Effect of Cyclically Pressurized Solid–Liquid Extraction Polyphenols from Cagnulari Grape Pomace on Oxidative Endothelial Cell Death"

Article Title: Protective Effect of Cyclically Pressurized Solid–Liquid Extraction Polyphenols from Cagnulari Grape Pomace on Oxidative Endothelial Cell Death

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

doi: 10.3390/molecules23092105

MALDI-TOF MS analysis. ( A ) Raw extract spectrum, confirming the presence of several anthocyanins, including malvidin, malvidin-3- O -glucoside, peodinin-3- O -glucoside, and malvidin-3-(6-acetyl)-glucoside. ( B ) Fraction 1 and ( C ) Fraction 2 spectra, confirming the presence of peodinin-3- O -glucoside and malvidin-3- O -glucoside, respectively.
Figure Legend Snippet: MALDI-TOF MS analysis. ( A ) Raw extract spectrum, confirming the presence of several anthocyanins, including malvidin, malvidin-3- O -glucoside, peodinin-3- O -glucoside, and malvidin-3-(6-acetyl)-glucoside. ( B ) Fraction 1 and ( C ) Fraction 2 spectra, confirming the presence of peodinin-3- O -glucoside and malvidin-3- O -glucoside, respectively.

Techniques Used: Mass Spectrometry

70) Product Images from "Earwax as an alternative specimen for forensic analysis"

Article Title: Earwax as an alternative specimen for forensic analysis

Journal: Forensic Toxicology

doi: 10.1007/s11419-017-0363-z

MRM chromatograms of actual earwax samples of a subject #1 showing peaks of oxcarbamazepine, levetiracetam, and topiramate; b subject #15 showing peaks of lamotrigine, clobazam, and phenobarbital; c subject #4 showing peaks of carbamazepine, phenytoin, and valproic acid; d subject #5 showing peaks of lacosamide; e subject #16 showing peak of clonazepam; f subject #17 showing peak of clozapine
Figure Legend Snippet: MRM chromatograms of actual earwax samples of a subject #1 showing peaks of oxcarbamazepine, levetiracetam, and topiramate; b subject #15 showing peaks of lamotrigine, clobazam, and phenobarbital; c subject #4 showing peaks of carbamazepine, phenytoin, and valproic acid; d subject #5 showing peaks of lacosamide; e subject #16 showing peak of clonazepam; f subject #17 showing peak of clozapine

Techniques Used:

71) Product Images from "A New Safety Concern for Glaucoma Treatment Demonstrated by Mass Spectrometry Imaging of Benzalkonium Chloride Distribution in the Eye, an Experimental Study in Rabbits"

Article Title: A New Safety Concern for Glaucoma Treatment Demonstrated by Mass Spectrometry Imaging of Benzalkonium Chloride Distribution in the Eye, an Experimental Study in Rabbits

Journal: PLoS ONE

doi: 10.1371/journal.pone.0050180

MALDI-TOF imaging of whole eye section of a control rabbit. MALDI-TOF imaging shows the absence of benzalkonium chloride (BAK) in the control eye. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), nasal iridocorneal angle (area 2) and near to the optic nerve (area 3). (b, c) Overlays between HE and MALDI-TOF images of BAK C 12 and C 14 eye distributions at m/z 304.32 and 332.36, respectively. Intensities of the ions are represented in colour, based on the intensity scale provided (from black to white). Field of view 18× 6 mm. (d, e, f) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively confirming the absence of BAK C 12 and C 14 .
Figure Legend Snippet: MALDI-TOF imaging of whole eye section of a control rabbit. MALDI-TOF imaging shows the absence of benzalkonium chloride (BAK) in the control eye. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), nasal iridocorneal angle (area 2) and near to the optic nerve (area 3). (b, c) Overlays between HE and MALDI-TOF images of BAK C 12 and C 14 eye distributions at m/z 304.32 and 332.36, respectively. Intensities of the ions are represented in colour, based on the intensity scale provided (from black to white). Field of view 18× 6 mm. (d, e, f) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively confirming the absence of BAK C 12 and C 14 .

Techniques Used: Imaging, Staining

Round-robin experiment using the 4800 MALDI-TOF/TOF mass spectrometer (ICSN-CNRS). MALDI-TOF imaging generated by the 4800 MALDI-TOF/TOF mass spectrometer (ICSN-CNRS) shows the BAK distribution in whole eye section of a rabbit instilled once a day with one drop of 0.2% benzalkonium chloride (BAK) for 1 month. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), near to optic nerve area (area 2) and optic nerve (area 3). (b, c) Overlays between HE staining and MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.32 and m/z 332.36, respectively. (d, e) MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and m/z 332.33, respectively, with intensity scale from 0 to 714. Intensities of the ions are represented in color, based on the intensity scale provided (from black to red). Field of view 18×23 mm. (f, g, h) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively, showing BAK C 12 and C 14 ion peaks.
Figure Legend Snippet: Round-robin experiment using the 4800 MALDI-TOF/TOF mass spectrometer (ICSN-CNRS). MALDI-TOF imaging generated by the 4800 MALDI-TOF/TOF mass spectrometer (ICSN-CNRS) shows the BAK distribution in whole eye section of a rabbit instilled once a day with one drop of 0.2% benzalkonium chloride (BAK) for 1 month. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), near to optic nerve area (area 2) and optic nerve (area 3). (b, c) Overlays between HE staining and MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.32 and m/z 332.36, respectively. (d, e) MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and m/z 332.33, respectively, with intensity scale from 0 to 714. Intensities of the ions are represented in color, based on the intensity scale provided (from black to red). Field of view 18×23 mm. (f, g, h) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively, showing BAK C 12 and C 14 ion peaks.

Techniques Used: Mass Spectrometry, Imaging, Generated, Staining

Chemical structures and MALDI-TOF spectra. Chemical structure and MALDI-TOF spectrum of benzalkonium homologs used in this study, BAK C 12 and BAK C 14 , are presented with their respective MALDI-TOF spectrum in positive ion mode. The BAK solution instilled in the rabbit eyes contained two thirds of BAK C 12 ( m/z 304.30) and one third of BAK C 14 ( m/z 332.33).
Figure Legend Snippet: Chemical structures and MALDI-TOF spectra. Chemical structure and MALDI-TOF spectrum of benzalkonium homologs used in this study, BAK C 12 and BAK C 14 , are presented with their respective MALDI-TOF spectrum in positive ion mode. The BAK solution instilled in the rabbit eyes contained two thirds of BAK C 12 ( m/z 304.30) and one third of BAK C 14 ( m/z 332.33).

Techniques Used:

MALDI-TOF imaging of whole eye section of a rabbit instilled twice a day with one drop of 0.01% benzalkonium chloride (BAK) for 5 months. MALDI-TOF imaging shows the BAK distribution in a BAK-treated eye. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), nasal iridocorneal angle (area 2) and optic nerve area (area 3). (b, c) Overlays between HE staining and MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and 332.33, respectively. (d, e) MALDI-TOF ion images of BAK C 12 and C 14 distributions at m/z 304.30 and 332.33, respectively. Intensities of the ions are represented in colour, based on the intensity scale provided (from black to white). Field of view 16×15 mm. (f, g, h) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively, showing BAK C 12 and C 14 ion peaks.
Figure Legend Snippet: MALDI-TOF imaging of whole eye section of a rabbit instilled twice a day with one drop of 0.01% benzalkonium chloride (BAK) for 5 months. MALDI-TOF imaging shows the BAK distribution in a BAK-treated eye. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), nasal iridocorneal angle (area 2) and optic nerve area (area 3). (b, c) Overlays between HE staining and MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and 332.33, respectively. (d, e) MALDI-TOF ion images of BAK C 12 and C 14 distributions at m/z 304.30 and 332.33, respectively. Intensities of the ions are represented in colour, based on the intensity scale provided (from black to white). Field of view 16×15 mm. (f, g, h) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively, showing BAK C 12 and C 14 ion peaks.

Techniques Used: Imaging, Staining

Round-robin experiment using the AutoFlex speed LRF MALDI-TOF mass spectrometer (ImaBiotech). MALDI-TOF imaging generated by the AutoFlex speed LRF MALDI-TOF mass spectrometer (ImaBiotech) shows the BAK distribution in whole eye section of a rabbit instilled once a day with one drop of 0.2% benzalkonium chloride (BAK) for 1 month. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), nasal iridocorneal angle (area 2) and optic nerve area (area 3). (b, c) Overlays between HE staining and MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and 332.33, respectively. (d, e) MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and m/z 332.33, respectively. Intensities of the ions are represented in colour, based on the intensity scale provided (from black to white). Field of view 8×10 mm. (f, g, h) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively, showing BAK C 12 and C 14 ion peaks.
Figure Legend Snippet: Round-robin experiment using the AutoFlex speed LRF MALDI-TOF mass spectrometer (ImaBiotech). MALDI-TOF imaging generated by the AutoFlex speed LRF MALDI-TOF mass spectrometer (ImaBiotech) shows the BAK distribution in whole eye section of a rabbit instilled once a day with one drop of 0.2% benzalkonium chloride (BAK) for 1 month. (a) Histology image of an adjacent cryosection stained with hematoxylin-eosin (HE) showing three areas of interest: cornea (area 1), nasal iridocorneal angle (area 2) and optic nerve area (area 3). (b, c) Overlays between HE staining and MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and 332.33, respectively. (d, e) MALDI-TOF ion images of BAK C 12 and C 14 distributions in whole eye section at m/z 304.30 and m/z 332.33, respectively. Intensities of the ions are represented in colour, based on the intensity scale provided (from black to white). Field of view 8×10 mm. (f, g, h) MALDI-TOF mass spectra extracted from areas 1, 2 and 3, respectively, showing BAK C 12 and C 14 ion peaks.

Techniques Used: Mass Spectrometry, Imaging, Generated, Staining

72) Product Images from "Accumulation of catechins and expression of catechin synthetic genes in Camellia sinensis at different developmental stages"

Article Title: Accumulation of catechins and expression of catechin synthetic genes in Camellia sinensis at different developmental stages

Journal: Botanical Studies

doi: 10.1186/s40529-016-0143-9

Chromatogram of the catechin components in tea. Results from a representative HPLC experiment are shown. The individual peaks represent A (gallocatechin; GC), B (epicatechin gallate; EGC), C (catechin; C), D (epicatechin; EC), E (epigallocatedchin gallate; EGCG), F (gallocatechin gallate; GCG), and G (epicatechin gallate; ECG)
Figure Legend Snippet: Chromatogram of the catechin components in tea. Results from a representative HPLC experiment are shown. The individual peaks represent A (gallocatechin; GC), B (epicatechin gallate; EGC), C (catechin; C), D (epicatechin; EC), E (epigallocatedchin gallate; EGCG), F (gallocatechin gallate; GCG), and G (epicatechin gallate; ECG)

Techniques Used: High Performance Liquid Chromatography

73) Product Images from "Increased Oral Bioavailability of Resveratrol by Its Encapsulation in Casein Nanoparticles"

Article Title: Increased Oral Bioavailability of Resveratrol by Its Encapsulation in Casein Nanoparticles

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19092816

Resveratrol release profile from casein nanoparticles after incubation in simulated gastric (SGI, pH 1.2; 0–2 h) and simulated intestinal fluids (SIF, pH 6.8; 2–24 h) under sink conditions. Data expressed as mean ± SD, n = 3.
Figure Legend Snippet: Resveratrol release profile from casein nanoparticles after incubation in simulated gastric (SGI, pH 1.2; 0–2 h) and simulated intestinal fluids (SIF, pH 6.8; 2–24 h) under sink conditions. Data expressed as mean ± SD, n = 3.

Techniques Used: Incubation

Scanning Electron Microscopy microphotographs obtained from resveratrol-loaded casein nanoparticles.
Figure Legend Snippet: Scanning Electron Microscopy microphotographs obtained from resveratrol-loaded casein nanoparticles.

Techniques Used: Electron Microscopy

Resveratrol plasma concentration vs. time after a single oral administration of 15 mg/kg for the different formulations tested. (i) Resveratrol PEG400:water solution (●); (ii) Resveratrol-loaded casein nanoparticles (■). Data expressed as mean ± SD, ( n = 6).
Figure Legend Snippet: Resveratrol plasma concentration vs. time after a single oral administration of 15 mg/kg for the different formulations tested. (i) Resveratrol PEG400:water solution (●); (ii) Resveratrol-loaded casein nanoparticles (■). Data expressed as mean ± SD, ( n = 6).

Techniques Used: Concentration Assay

Resveratrol- O -3-glucuronide plasma concentration vs time after the administration of a single dose of resveratrol (15 mg/kg) in casein nanoparticles (oral) or dissolved in a PEG400:water mixture (oral and intravenously). (i) Resveratrol solution intravenously administered (Rsv-IV) (●); (ii) resveratrol solution orally administered (Rsv-Sol) (■); (iii) resveratrol-loaded casein nanoparticles orally administered (Rsv-NP-C) (▲). Data expressed as mean ± SD, ( n = 6).
Figure Legend Snippet: Resveratrol- O -3-glucuronide plasma concentration vs time after the administration of a single dose of resveratrol (15 mg/kg) in casein nanoparticles (oral) or dissolved in a PEG400:water mixture (oral and intravenously). (i) Resveratrol solution intravenously administered (Rsv-IV) (●); (ii) resveratrol solution orally administered (Rsv-Sol) (■); (iii) resveratrol-loaded casein nanoparticles orally administered (Rsv-NP-C) (▲). Data expressed as mean ± SD, ( n = 6).

Techniques Used: Concentration Assay

In vitro-in vivo correlation (IVIVC) plot for resveratrol when loaded in casein nanoparticles (Rsv-NP-C), showing fraction absorbed as a function of fraction released.
Figure Legend Snippet: In vitro-in vivo correlation (IVIVC) plot for resveratrol when loaded in casein nanoparticles (Rsv-NP-C), showing fraction absorbed as a function of fraction released.

Techniques Used: In Vitro, In Vivo

74) Product Images from "Metabolic Engineering for Resveratrol Derivative Biosynthesis in Escherichia coli"

Article Title: Metabolic Engineering for Resveratrol Derivative Biosynthesis in Escherichia coli

Journal: Molecules and Cells

doi: 10.14348/molcells.2015.2188

(A) Biosynthetic pathway of stilbene compound production from phenylalanine and (B) construction of recombinant plasmids carrying the genes ( ScCCL , STS , the synthetic RpSTSsyn , VrROMTsyn , and SbROMT3syn ) involved in stilbene biosynthesis. The sequential actions of PAL or TAL, C4H, 4CL (CCL), STS, and ROMT result in the conversion of phenylalanine to stilbenes, resveratrol, and its methylated derivatives. PAL, phenylalanine ammonia-lyase; C4H, cinnamate-4-hydroxylase; TAL, tyrosine ammonia-lyase; 4CL, 4-coumarate:coenzyme A ligase; CCL, cinnamate/4-coumarate:coenzyme A ligase; STS, stilbene synthase; ROMT, resveratrol O -methyltransferase; T7 Pro, T7 RNA polymerase promoter; H, His-tag; S, S-tag.
Figure Legend Snippet: (A) Biosynthetic pathway of stilbene compound production from phenylalanine and (B) construction of recombinant plasmids carrying the genes ( ScCCL , STS , the synthetic RpSTSsyn , VrROMTsyn , and SbROMT3syn ) involved in stilbene biosynthesis. The sequential actions of PAL or TAL, C4H, 4CL (CCL), STS, and ROMT result in the conversion of phenylalanine to stilbenes, resveratrol, and its methylated derivatives. PAL, phenylalanine ammonia-lyase; C4H, cinnamate-4-hydroxylase; TAL, tyrosine ammonia-lyase; 4CL, 4-coumarate:coenzyme A ligase; CCL, cinnamate/4-coumarate:coenzyme A ligase; STS, stilbene synthase; ROMT, resveratrol O -methyltransferase; T7 Pro, T7 RNA polymerase promoter; H, His-tag; S, S-tag.

Techniques Used: Recombinant, Methylation

Time-course production of resveratrol and pinostilbene in recombinant E. coli . E. coli cells harboring CCL+STS, CCL+STS-VrROMT, or CCL+STS-SbROMT3 construct were cultured and harvested at the indicated time points after the addition of 1 mM p- coumaric acid to the pre-induced cells. E. coli cells carrying both pCOLADuet-1 and pETDuet-1 were used as a control. Samples were subjected to HPLC analysis. The amounts (mg/L) of resveratrol (A) and pinostilbene (B) at the indicated time points were measured by comparison with standard curves generated from known concentrations of resveratrol and pinostilbene.
Figure Legend Snippet: Time-course production of resveratrol and pinostilbene in recombinant E. coli . E. coli cells harboring CCL+STS, CCL+STS-VrROMT, or CCL+STS-SbROMT3 construct were cultured and harvested at the indicated time points after the addition of 1 mM p- coumaric acid to the pre-induced cells. E. coli cells carrying both pCOLADuet-1 and pETDuet-1 were used as a control. Samples were subjected to HPLC analysis. The amounts (mg/L) of resveratrol (A) and pinostilbene (B) at the indicated time points were measured by comparison with standard curves generated from known concentrations of resveratrol and pinostilbene.

Techniques Used: Recombinant, Construct, Cell Culture, High Performance Liquid Chromatography, Generated

LC-MS analysis of resveratrol, pinostilbene and pterostilbene produced by recombinant E. coli cells. The molecular masses of resveratrol, pinostilbene and pterostilbene were analyzed with a linear ion trap quadrupole LC-MS at a positive mode of atmospheric pressure chemical ionization. Samples were taken from cell culture at 48 h after the addition of p -coumaric acid, and the extracted compounds were analyzed by LC-MS. The injection volume was 10 μl. Chromatograms (A), (B) and (C) represent the LC-MS spectra showing the peaks at m/z 229.2, 243.1 and 257.1 for reveratrol, pinostilbene and pterostilbene, respectively.
Figure Legend Snippet: LC-MS analysis of resveratrol, pinostilbene and pterostilbene produced by recombinant E. coli cells. The molecular masses of resveratrol, pinostilbene and pterostilbene were analyzed with a linear ion trap quadrupole LC-MS at a positive mode of atmospheric pressure chemical ionization. Samples were taken from cell culture at 48 h after the addition of p -coumaric acid, and the extracted compounds were analyzed by LC-MS. The injection volume was 10 μl. Chromatograms (A), (B) and (C) represent the LC-MS spectra showing the peaks at m/z 229.2, 243.1 and 257.1 for reveratrol, pinostilbene and pterostilbene, respectively.

Techniques Used: Liquid Chromatography with Mass Spectroscopy, Produced, Recombinant, Cell Culture, Injection

HPLC analysis of resveratrol and pinostilbene produced by recombinant E. coli . E. coli cells harboring CCL+STS, CCL+STS-VrROMT, or CCL+STS-SbROMT3 constructs were cultured for 48 h prior to extraction. Chromatogram STD represents the authentic standards of resveratrol (Res), pinostilbene (Pino), and pterostilbene (Ptero) with retention times of 9.423, 19.327, and 32.009 min, respectively. Chromatograms Control, CCL+STS, CCL+STS-VrROMT, and CCL+STS-SbROMT3 represent the samples produced from p- coumaric acid by recombinant E. coli . Insets indicate the chromatograms magnified to show small amounts of pterostilbene production.
Figure Legend Snippet: HPLC analysis of resveratrol and pinostilbene produced by recombinant E. coli . E. coli cells harboring CCL+STS, CCL+STS-VrROMT, or CCL+STS-SbROMT3 constructs were cultured for 48 h prior to extraction. Chromatogram STD represents the authentic standards of resveratrol (Res), pinostilbene (Pino), and pterostilbene (Ptero) with retention times of 9.423, 19.327, and 32.009 min, respectively. Chromatograms Control, CCL+STS, CCL+STS-VrROMT, and CCL+STS-SbROMT3 represent the samples produced from p- coumaric acid by recombinant E. coli . Insets indicate the chromatograms magnified to show small amounts of pterostilbene production.

Techniques Used: High Performance Liquid Chromatography, Produced, Recombinant, Construct, Cell Culture

Expression of His-tagged plant STS and ScCCL recombinant proteins and time-course production of resveratrol in E. coli cells. (A) E. coli cells harboring both pCOLADuet-H::CCL and pET22b-STS::H (CCL+STS) constructs were grown in modified M9 media and induced by the addition of 0.5 mM IPTG at 25°C for 2 h and 4 h. Expression of His-tag fusion proteins was confirmed by Western blot analysis with anti-His-tag antibody. M, protein molecular marker in kDa; S, soluble fraction (20 μg); P, insoluble pellet fraction (1 μg). Arrows and arrowheads indicate the expressed ScCCL and plant STS recombinant proteins, respectively. (B) E. coli cells harboring CCL+STS constructs were cultured and harvested at the indicated time points after the addition of 1 mM p -coumaric acid to the pre-induced cells. E. coli cells carrying both pCOLADuet-1 and pET-22b were used as a control. Samples were extracted with ethyl acetate and subjected to HPLC analysis. E. coli cells were pre-induced to express recombinant proteins by the addition of 0.1 mM IPTG. (C) HPLC analysis was performed using a C18 reverse-phase column. Chromatogram STD represents the authentic standards of resveratrol (Res) and p -coumaric acid ( p -Cou) with retention times of 9.887 and 4.907 min, respectively.
Figure Legend Snippet: Expression of His-tagged plant STS and ScCCL recombinant proteins and time-course production of resveratrol in E. coli cells. (A) E. coli cells harboring both pCOLADuet-H::CCL and pET22b-STS::H (CCL+STS) constructs were grown in modified M9 media and induced by the addition of 0.5 mM IPTG at 25°C for 2 h and 4 h. Expression of His-tag fusion proteins was confirmed by Western blot analysis with anti-His-tag antibody. M, protein molecular marker in kDa; S, soluble fraction (20 μg); P, insoluble pellet fraction (1 μg). Arrows and arrowheads indicate the expressed ScCCL and plant STS recombinant proteins, respectively. (B) E. coli cells harboring CCL+STS constructs were cultured and harvested at the indicated time points after the addition of 1 mM p -coumaric acid to the pre-induced cells. E. coli cells carrying both pCOLADuet-1 and pET-22b were used as a control. Samples were extracted with ethyl acetate and subjected to HPLC analysis. E. coli cells were pre-induced to express recombinant proteins by the addition of 0.1 mM IPTG. (C) HPLC analysis was performed using a C18 reverse-phase column. Chromatogram STD represents the authentic standards of resveratrol (Res) and p -coumaric acid ( p -Cou) with retention times of 9.887 and 4.907 min, respectively.

Techniques Used: Expressing, Recombinant, Construct, Modification, Western Blot, Marker, Cell Culture, Positron Emission Tomography, High Performance Liquid Chromatography

In vitro assays for production of resveratrol from p -coumaric acid by purified recombinant CCL and STS. His-tagged CCL (A) and STS (B) recombinant proteins were expressed in E. coli and affinity-purified using Ni-NTA agarose. Protein samples (2 μg) of total lysates (T) after induction, unbound fraction (U) after Ni-NTA bead incubation, and purified His-CCL or His-STS eluted from agarose beads (E) were separated by 13% SDS-PAGE and stained with CBB. The partially purified enzymes (each 1.5 μM) were assayed for CCL and STS activity with 0.2 mM p -coumaric acid as a precursor in the presence of 0.3 mM malonyl-CoA and 0.3 mM CoA. The reaction mixtures were incubated at 30°C for 3 h prior to extraction. Concentrated 20 μl samples were analyzed by HPLC. Chromatogram STD represents the authentic standards of p -coumaric acid ( p -Cou) and resveratrol (Res) with retention times of 4.75 and 9.51 min, respectively. Chromatograms Control and CCL+ STS represent the samples produced from p -coumaric acid by in vitro assays without and with recombinant proteins, respectively.
Figure Legend Snippet: In vitro assays for production of resveratrol from p -coumaric acid by purified recombinant CCL and STS. His-tagged CCL (A) and STS (B) recombinant proteins were expressed in E. coli and affinity-purified using Ni-NTA agarose. Protein samples (2 μg) of total lysates (T) after induction, unbound fraction (U) after Ni-NTA bead incubation, and purified His-CCL or His-STS eluted from agarose beads (E) were separated by 13% SDS-PAGE and stained with CBB. The partially purified enzymes (each 1.5 μM) were assayed for CCL and STS activity with 0.2 mM p -coumaric acid as a precursor in the presence of 0.3 mM malonyl-CoA and 0.3 mM CoA. The reaction mixtures were incubated at 30°C for 3 h prior to extraction. Concentrated 20 μl samples were analyzed by HPLC. Chromatogram STD represents the authentic standards of p -coumaric acid ( p -Cou) and resveratrol (Res) with retention times of 4.75 and 9.51 min, respectively. Chromatograms Control and CCL+ STS represent the samples produced from p -coumaric acid by in vitro assays without and with recombinant proteins, respectively.

Techniques Used: In Vitro, Purification, Recombinant, Affinity Purification, Incubation, SDS Page, Staining, Activity Assay, High Performance Liquid Chromatography, Produced

75) Product Images from "Simultaneous Analysis of Secondary Structure and Light Scattering from Circular Dichroism Titrations: Application to Vectofusin-1"

Article Title: Simultaneous Analysis of Secondary Structure and Light Scattering from Circular Dichroism Titrations: Application to Vectofusin-1

Journal: Scientific Reports

doi: 10.1038/srep39450

Comparison between light scattering and PCA decomposition of CD spectra. ( A ) Static and dynamic light scattering. The derived count rate (DCR) is a measure for the amount of backscattered light and is compared between a control curve of liposomes in buffer (black crosses) and a titration of vectofusin-1 with the same liposomes (open black circles). The estimated diameter of the particle calculated from dynamic light scattering is shown as the z-average in the same figure on the scondary axis (solid black circles), where the first datapoint is left out because the low scattering of peptide alone makes the computation of the z-average unreliable. ( B ) PCA scores as function of lipid/peptide ratio.
Figure Legend Snippet: Comparison between light scattering and PCA decomposition of CD spectra. ( A ) Static and dynamic light scattering. The derived count rate (DCR) is a measure for the amount of backscattered light and is compared between a control curve of liposomes in buffer (black crosses) and a titration of vectofusin-1 with the same liposomes (open black circles). The estimated diameter of the particle calculated from dynamic light scattering is shown as the z-average in the same figure on the scondary axis (solid black circles), where the first datapoint is left out because the low scattering of peptide alone makes the computation of the z-average unreliable. ( B ) PCA scores as function of lipid/peptide ratio.

Techniques Used: Derivative Assay, Titration

( A ) Titration of vectofusin-1 with POPC/POPS liposomes. Spectra are corrected with control curves of liposomes only but peptide-induced liposome aggregation causes some of the spectra to be severly affected by light scattering. ( B ) Spectral components as determined by principal component analysis. The CD spectra are fully described by two components that we interpret as light scattering (PC1) and peptide structural changes (PC2).
Figure Legend Snippet: ( A ) Titration of vectofusin-1 with POPC/POPS liposomes. Spectra are corrected with control curves of liposomes only but peptide-induced liposome aggregation causes some of the spectra to be severly affected by light scattering. ( B ) Spectral components as determined by principal component analysis. The CD spectra are fully described by two components that we interpret as light scattering (PC1) and peptide structural changes (PC2).

Techniques Used: Titration

Related Articles

Cell Cycle Assay:

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Cytometry:

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Incubation:

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Activity Assay:

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High Performance Liquid Chromatography:

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Transfection:

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Activation Assay:

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Cell Culture:

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Inhibition:

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CtB Assay:

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Recombinant:

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MTT Assay:

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Methylation:

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Isolation:

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Flow Cytometry:

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Labeling:

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Mouse Assay:

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Small Interfering RNA:

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Software:

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Quantitation Assay:

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Concentration Assay:

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Marker:

Article Title: Human Immunodeficiency Virus Type 1 Enters Primary Human Brain Microvascular Endothelial Cells by a Mechanism Involving Cell Surface Proteoglycans Independent of Lipid Rafts
Article Snippet: Anti-human von Willebrand factor antibody was purchased from Sigma (St. Louis, Mo.), anti-ZO-1 antibody was purchased from Zymed (South San Francisco, Calif.), anti-heparin/heparan sulfate (recognizes intact heparan sulfates) antibody was from Research Diagnostics, Inc. (Flanders, N.J.), and anti-chondroitin sulfate (recognizes intact chondroitin-6-sulfate and/or chondroitin-4-sulfates) antibody and the fluorescent conjugate (Alexa Fluor 488) of cholera toxin subunit B (CTB), which is a marker for ganglioside GM1-containing lipid rafts, were obtained from Molecular Probes Inc. (Eugene, Oreg.). .. For cholesterol depletion, we used randomly methylated β-cyclodextrin (BCD) (commercial name, TRMBP) and hydroxypropyl BCD (commercial name, THPBP) from Cyclodextrin Technologies Development, Inc. (High Springs, Fla.), and for inhibition of cholesterol synthesis, we used the 3-hydroxy-3-methylglutaryl (HMG)-coenzyme A (CoA) reductase inhibitors mevinolin (Sigma), atorvastatin, and pravastatin (Calbiochem, San Diego, Calif.).

other:

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Article Snippet: Reagents Compounds used are T0901317 (Sigma, T2320) GW501516 (Enzo life sciences, 89150-762), Rosiglitazone (Sigma, R2408), Atorvastatin calcium salt (Sigma, PZ001), Simvastatin (Sigma, S6196), Rosuvastatin (Sigma, SML1264), Fluvastatin (Sigma, SML0038), Mevastatin (Tocris, 1526) Lovastatin (Sigma, PHR1285), Mevalonolactone (Mevalonic Acid) (Sigma, M4777), (R)-Mevalonic acid 5-pyrophosphoate tetralithium salt (Sigma, 77631), Mevalonic acid 5-phosphate thrilithium salt hydrate (Sigma, 79849), Methyl-β-cyclodextrin (Sigma, C4555), YM-53601 (Cayman Chemicals, 18113), AY 9944 dihydrochloride (Tocris), FTI-227 trifluoroacetate salt (Sigma, F9803), GGTI-298 trifluoracetate salt hydrate (Sigma, G5169), Fatostatin hydrobromide (Sigma, F8932), 24(S)-hydroxycholesterol (Cayman Chemicals, 10009931), K-604 (Sigma-Aldrich, SML1837), chloroquine diphosphate salt (Sigma, C6628), MG-132 (InSolution, Calbiochem, 37391), Cholestane (Sigma-Aldrich C8003) N -Methyl-N -(trimethylsilyl)trifluoroacetamide (MSTFA, ThermoFisher TS48910), 1 μL of dry pyridine (Sigma-Aldrich, 270970) and sodium hydroxide solution 10M (BioUltra, Sigma Aldrich, 72068).

Article Title: Punicalagin Decreases Serum Glucose Levels and Increases PON1 Activity and HDL Anti-Inflammatory Values in Balb/c Mice Fed a High-Fat Diet
Article Snippet: Punicalagin, quercetin, Tween 80, methanol, chloroform, atorvastatin, dihydrocoumarin, dimethyl sulfoxide (DMSO), 1-palmitoyl-2-arachidonoyl- sn -glycero-3-phosphorylcholine (PAPC), and dichloro-dihydro-fluorescein diacetate (DCFH-DA) were purchased from Sigma-Aldrich; 1-palmitoyl-2-(5,6-epoxyisoprostane E2)- sn -glycero-3-phosphocholine (PEIPC) was prepared from PAPC, and ALZET osmotic minipumps were purchased from Biotest.

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  • 85
    Millipore imatinib hydrochloride salt
    The comparison of time–activity curves of the (A) heart, (B) lungs, (C) kidneys, (D) spleen, (E) liver and (F) gallbladder before and after pretreatment with 32 mg of <t>imatinib</t> administration by intravenous injection. Time–activity curves
    Imatinib Hydrochloride Salt, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore jnk inhibitor v
    POH-induced apoptosis in U251 cells. U251 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM <t>JNK</t> <t>inhibitor</t> V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P
    Jnk Inhibitor V, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Millipore as 601245
    POH-induced apoptosis in U251 cells. U251 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM <t>JNK</t> <t>inhibitor</t> V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P
    As 601245, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore acetonitrile water
    POH-induced apoptosis in U251 cells. U251 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM <t>JNK</t> <t>inhibitor</t> V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P
    Acetonitrile Water, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The comparison of time–activity curves of the (A) heart, (B) lungs, (C) kidneys, (D) spleen, (E) liver and (F) gallbladder before and after pretreatment with 32 mg of imatinib administration by intravenous injection. Time–activity curves

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: The comparison of time–activity curves of the (A) heart, (B) lungs, (C) kidneys, (D) spleen, (E) liver and (F) gallbladder before and after pretreatment with 32 mg of imatinib administration by intravenous injection. Time–activity curves

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques: Activity Assay, Injection

    Structure of imatinib (the active ingredient of Gleevec).

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: Structure of imatinib (the active ingredient of Gleevec).

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques:

    Synthetic scheme of norimatinib, imatinib and [ N - 11 C-methyl]imatinib.

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: Synthetic scheme of norimatinib, imatinib and [ N - 11 C-methyl]imatinib.

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques:

    PET image of baboon brain with [ N - 11 C-methyl]imatinib. Summed frames over 90 min after the injection of 3.9 mCi of [ N - 11 ]. The anesthesia

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: PET image of baboon brain with [ N - 11 C-methyl]imatinib. Summed frames over 90 min after the injection of 3.9 mCi of [ N - 11 ]. The anesthesia

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques: Positron Emission Tomography, Injection

    PET image of the torso of the anesthetized baboon with [ N - 11 C-methyl]imatinib. PET images (summation of frames over 90 min) of the baboon torso after the injection of 4.71 mCi of [ N - 11 C-methyl]imatinib showing the accumulation of carbon-11 in the heart

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: PET image of the torso of the anesthetized baboon with [ N - 11 C-methyl]imatinib. PET images (summation of frames over 90 min) of the baboon torso after the injection of 4.71 mCi of [ N - 11 C-methyl]imatinib showing the accumulation of carbon-11 in the heart

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques: Positron Emission Tomography, Injection

    (A) Time–activity curve at baseline and after imatinib pretreatment for [ N - 11 C-methyl]imatinib in plasma showing the first 5 min and (B) the plasma integral over 90 min. Pretreatment delayed [ N - 11 C-methyl]imatinib peak and reduced its accumulated

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: (A) Time–activity curve at baseline and after imatinib pretreatment for [ N - 11 C-methyl]imatinib in plasma showing the first 5 min and (B) the plasma integral over 90 min. Pretreatment delayed [ N - 11 C-methyl]imatinib peak and reduced its accumulated

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques: Activity Assay

    Time–activity curves for two different anesthetized baboons (A and B) that each received two injections of [ N - 11 C-methyl]imatinib in the brain through the urinary bladder.

    Journal: Nuclear medicine and biology

    Article Title: Synthesis and positron emission tomography studies of carbon-11-labeled imatinib (Gleevec)

    doi: 10.1016/j.nucmedbio.2006.11.004

    Figure Lengend Snippet: Time–activity curves for two different anesthetized baboons (A and B) that each received two injections of [ N - 11 C-methyl]imatinib in the brain through the urinary bladder.

    Article Snippet: The excess amount of hydrochloric acid was removed by coevaporation with acetonitrile to give imatinib hydrochloride salt, which was dissolved in 5 ml of sterile water and passed through a Millipore filter.

    Techniques: Activity Assay

    POH-induced apoptosis in U251 cells. U251 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM JNK inhibitor V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P

    Journal: Molecular Cancer

    Article Title: Na/K-ATPase as a target for anticancer drugs: studies with perillyl alcohol

    doi: 10.1186/s12943-015-0374-5

    Figure Lengend Snippet: POH-induced apoptosis in U251 cells. U251 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM JNK inhibitor V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P

    Article Snippet: Cell death assay U87 and U251 cells were pretreated for 30 minutes with JNK inhibitor V [1,3-Benzothiazol-2-yl-(2-((2-(3-pyridinyl)ethyl)amino)-4-pyrimidinyl)acetonitrile; Calbiochem], an inhibitor of JNK1/2 activation, before treatment with 0.5 mM POH and 0.5 mM POH plus 0.5 μM JNK inhibitor V. After 24 hours of incubation, the cells were suspended in annexin and propidium iodide binding buffer as specified in the TACS Annexin V-FITC apoptosis detection kit (R & D Systems).

    Techniques:

    The effects of JNK inhibition on the induction of cell death by POH in U251 cells. Before treatment, U251 cells were incubated without (A and B) or with (C and D) JNK inhibitor V (0.5 μM) for 30 minutes. The cells were treated with 0.1% DMSO (A) , 0.5 mM POH (B) 0.1% DMSO plus JNK inhibitor V (C) , and 0.5 mM POH plus JNK inhibitor V (D) . After 24 hours of incubation, the cells were stained with annexin V-FITC and propidium iodide and analyzed by flow cytometry. Figure 7E represents the percentage of dead cells as indicated by early apoptosis and late apoptosis or necrosis (right lower quadrant + right upper quadrant, respectively), which was calculated from the data shown in Figures 7A-D. The data were expressed as the means ± SD from at least three different experiments. ***p

    Journal: Molecular Cancer

    Article Title: Na/K-ATPase as a target for anticancer drugs: studies with perillyl alcohol

    doi: 10.1186/s12943-015-0374-5

    Figure Lengend Snippet: The effects of JNK inhibition on the induction of cell death by POH in U251 cells. Before treatment, U251 cells were incubated without (A and B) or with (C and D) JNK inhibitor V (0.5 μM) for 30 minutes. The cells were treated with 0.1% DMSO (A) , 0.5 mM POH (B) 0.1% DMSO plus JNK inhibitor V (C) , and 0.5 mM POH plus JNK inhibitor V (D) . After 24 hours of incubation, the cells were stained with annexin V-FITC and propidium iodide and analyzed by flow cytometry. Figure 7E represents the percentage of dead cells as indicated by early apoptosis and late apoptosis or necrosis (right lower quadrant + right upper quadrant, respectively), which was calculated from the data shown in Figures 7A-D. The data were expressed as the means ± SD from at least three different experiments. ***p

    Article Snippet: Cell death assay U87 and U251 cells were pretreated for 30 minutes with JNK inhibitor V [1,3-Benzothiazol-2-yl-(2-((2-(3-pyridinyl)ethyl)amino)-4-pyrimidinyl)acetonitrile; Calbiochem], an inhibitor of JNK1/2 activation, before treatment with 0.5 mM POH and 0.5 mM POH plus 0.5 μM JNK inhibitor V. After 24 hours of incubation, the cells were suspended in annexin and propidium iodide binding buffer as specified in the TACS Annexin V-FITC apoptosis detection kit (R & D Systems).

    Techniques: Inhibition, Incubation, Staining, Flow Cytometry, Cytometry

    POH-induced apoptosis in U87 cells. U87 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM JNK inhibitor V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P

    Journal: Molecular Cancer

    Article Title: Na/K-ATPase as a target for anticancer drugs: studies with perillyl alcohol

    doi: 10.1186/s12943-015-0374-5

    Figure Lengend Snippet: POH-induced apoptosis in U87 cells. U87 in the control condition (Control, A) . The cells were treated with 0.5 mM POH (POH, B) and 0.5 mM POH plus 0.5 μM JNK inhibitor V (POH + IJNK, C) . After 24 hours, the cells were immunostained for cleaved caspase-3 and the number of positive cells was analyzed (D) . Whereas POH induced cell apoptosis, the addition of the JNK inhibitor completely inhibited this effect. The addition of DMSO or JNK inhibitor V alone had no effect on cell death. Scale bar: 20 μm.*P

    Article Snippet: Cell death assay U87 and U251 cells were pretreated for 30 minutes with JNK inhibitor V [1,3-Benzothiazol-2-yl-(2-((2-(3-pyridinyl)ethyl)amino)-4-pyrimidinyl)acetonitrile; Calbiochem], an inhibitor of JNK1/2 activation, before treatment with 0.5 mM POH and 0.5 mM POH plus 0.5 μM JNK inhibitor V. After 24 hours of incubation, the cells were suspended in annexin and propidium iodide binding buffer as specified in the TACS Annexin V-FITC apoptosis detection kit (R & D Systems).

    Techniques:

    The effects of JNK inhibition on the induction of cell death by POH in U87 cells. Before treatment, U87 cells were incubated without (A and B) or with (C and D) JNK inhibitor V (0.5 μM) for 30 minutes. The cells were treated with 0.1% DMSO (A), 0.5 mM POH (B) 0.1% DMSO plus JNK inhibitor V (C) , and 0.5 mM POH plus JNK inhibitor V (D) . After 24 hours of incubation, the cells were stained with annexin V-FITC and propidium iodide and analyzed by flow cytometry. Figure 6E represents the percentage of dead cells indicated by early apoptosis and late apoptosis or necrosis (right lower quadrant + right upper quadrant, respectively), which was calculated from the data shown in Figures 6A-D. The data were expressed as the means ± SD from at least three different experiments. ***p

    Journal: Molecular Cancer

    Article Title: Na/K-ATPase as a target for anticancer drugs: studies with perillyl alcohol

    doi: 10.1186/s12943-015-0374-5

    Figure Lengend Snippet: The effects of JNK inhibition on the induction of cell death by POH in U87 cells. Before treatment, U87 cells were incubated without (A and B) or with (C and D) JNK inhibitor V (0.5 μM) for 30 minutes. The cells were treated with 0.1% DMSO (A), 0.5 mM POH (B) 0.1% DMSO plus JNK inhibitor V (C) , and 0.5 mM POH plus JNK inhibitor V (D) . After 24 hours of incubation, the cells were stained with annexin V-FITC and propidium iodide and analyzed by flow cytometry. Figure 6E represents the percentage of dead cells indicated by early apoptosis and late apoptosis or necrosis (right lower quadrant + right upper quadrant, respectively), which was calculated from the data shown in Figures 6A-D. The data were expressed as the means ± SD from at least three different experiments. ***p

    Article Snippet: Cell death assay U87 and U251 cells were pretreated for 30 minutes with JNK inhibitor V [1,3-Benzothiazol-2-yl-(2-((2-(3-pyridinyl)ethyl)amino)-4-pyrimidinyl)acetonitrile; Calbiochem], an inhibitor of JNK1/2 activation, before treatment with 0.5 mM POH and 0.5 mM POH plus 0.5 μM JNK inhibitor V. After 24 hours of incubation, the cells were suspended in annexin and propidium iodide binding buffer as specified in the TACS Annexin V-FITC apoptosis detection kit (R & D Systems).

    Techniques: Inhibition, Incubation, Staining, Flow Cytometry, Cytometry