paclitaxel  (Millipore)


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    Name:
    Paclitaxel
    Description:
    Chemical structure taxoide
    Catalog Number:
    t7191
    Price:
    None
    Applications:
    Paclitaxel has been used to study its effects on tumor regression in mouse models of pancreatic ductal adenocarcinoma. Paclitaxel has also been used as an internal standard for chromatographic assays of docetaxel. Furthermore, paclitaxel has been used to analyze its effects on Caenorhabditis elegans embryos.
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    Millipore paclitaxel
    Paclitaxel
    Chemical structure taxoide
    https://www.bioz.com/result/paclitaxel/product/Millipore
    Average 99 stars, based on 4296 article reviews
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    paclitaxel - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "Reversing multidrug resistance in breast cancer cells by silencing ABC transporter genes with nanoparticle-facilitated delivery of target siRNAs"

    Article Title: Reversing multidrug resistance in breast cancer cells by silencing ABC transporter genes with nanoparticle-facilitated delivery of target siRNAs

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S30500

    Combination effect of two siRNAs targeting ABCG2 and ABCB1 (ABCG2_v1 and ABCB1_v4 respectively) co-delivered with carbonate apatite nanoparticles on MCF-7 cell viability in presence of traditionally used chemotherapeutic agents. Anti-ABC siRNAs-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCG2 (1 nM) and anti-ABCB1 siRNA (1 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 1 nM drug (doxorubicin, paclitaxel or cisplatin). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.
    Figure Legend Snippet: Combination effect of two siRNAs targeting ABCG2 and ABCB1 (ABCG2_v1 and ABCB1_v4 respectively) co-delivered with carbonate apatite nanoparticles on MCF-7 cell viability in presence of traditionally used chemotherapeutic agents. Anti-ABC siRNAs-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCG2 (1 nM) and anti-ABCB1 siRNA (1 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 1 nM drug (doxorubicin, paclitaxel or cisplatin). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.

    Techniques Used: Generated, Incubation, Transfection, MTT Assay, Small Interfering RNA

    Effect of carbonate apatite-mediated delivery of ABCG2-targeted siRNA (ABCG2_v1) on MCF-7 cell viability in presence of low dose of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCG2 siRNA (1 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 10 pM, 100 pM or 1 nM of doxorubicin ( A ) or paclitaxel ( B ) or 100 pM, 1 nM, 10 nM cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.
    Figure Legend Snippet: Effect of carbonate apatite-mediated delivery of ABCG2-targeted siRNA (ABCG2_v1) on MCF-7 cell viability in presence of low dose of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCG2 siRNA (1 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 10 pM, 100 pM or 1 nM of doxorubicin ( A ) or paclitaxel ( B ) or 100 pM, 1 nM, 10 nM cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.

    Techniques Used: Generated, Incubation, Transfection, MTT Assay, Small Interfering RNA

    Effect of carbonate apatite-mediated delivery of ABCG2-targeted siRNA (ABCG2_v1) on MCF-7 cell viability in presence of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCG2 siRNA (1 or 10 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 100 nM of doxorubicin ( A ) or paclitaxel ( B ) or cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.
    Figure Legend Snippet: Effect of carbonate apatite-mediated delivery of ABCG2-targeted siRNA (ABCG2_v1) on MCF-7 cell viability in presence of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCG2 siRNA (1 or 10 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 100 nM of doxorubicin ( A ) or paclitaxel ( B ) or cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.

    Techniques Used: Generated, Incubation, Transfection, MTT Assay, Small Interfering RNA

    Effect of carbonate apatite-mediated delivery of ABCB1-targeted siRNA (ABCB1_v4) on MCF-7 cell viability in presence of low dose of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCB1 siRNA (1 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 10 pM, 100 pM or 1 nM of doxorubicin ( A ) or paclitaxel ( B ) or 100 pM, 1 nM, 10 nM cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.
    Figure Legend Snippet: Effect of carbonate apatite-mediated delivery of ABCB1-targeted siRNA (ABCB1_v4) on MCF-7 cell viability in presence of low dose of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCB1 siRNA (1 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 10 pM, 100 pM or 1 nM of doxorubicin ( A ) or paclitaxel ( B ) or 100 pM, 1 nM, 10 nM cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.

    Techniques Used: Generated, Incubation, Transfection, MTT Assay, Small Interfering RNA

    Effect of carbonate apatite-mediated delivery of ABCB1-targeted siRNA (ABCB1_v4) on MCF-7 cell viability in presence of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCB1 siRNA (1 or 10 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 100 nM of doxorubicin ( A ) or paclitaxel ( B ) or cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.
    Figure Legend Snippet: Effect of carbonate apatite-mediated delivery of ABCB1-targeted siRNA (ABCB1_v4) on MCF-7 cell viability in presence of traditionally used chemotherapeutic agents. Anti-ABC siRNA-carbonate apatite complexes were generated by mixing exogenously added 3 mM calcium chloride in 1 mL bicarbonate-buffered DMEM (pH 7.4), followed by addition of anti-ABCB1 siRNA (1 or 10 nM) and incubation at 37°C for 30 minutes. Supplementation of 10% FBS was followed by addition of 100 nM of doxorubicin ( A ) or paclitaxel ( B ) or cisplatin ( C ). Note: Transfection of MCF-7 cells was performed with the siRNA/nanoparticle complexes in presence of the free drugs for a consecutive period of 48 hours and viability of the cells was determined using MTT assay. Abbreviations: NP, nanoparticles; siRNA, small interfering RNA; DOX, doxorubicin; PAC, paclitaxel; CISP, cisplatin.

    Techniques Used: Generated, Incubation, Transfection, MTT Assay, Small Interfering RNA

    2) Product Images from "Codeine induces human mast cell chemokine and cytokine production: involvement of G-protein activation"

    Article Title: Codeine induces human mast cell chemokine and cytokine production: involvement of G-protein activation

    Journal:

    doi: 10.1111/j.1398-9995.2007.01345.x

    Codeine-induced mast cell degranulation is partially calcium and glucocorticoid sensitive. (A) LAD2 were stimulated with codeine (2.5 μg/ml) in Ca 2+ -free or Ca 2+ -containing (1.8 mM) media for 30 min and β-hexosaminidase release was measured
    Figure Legend Snippet: Codeine-induced mast cell degranulation is partially calcium and glucocorticoid sensitive. (A) LAD2 were stimulated with codeine (2.5 μg/ml) in Ca 2+ -free or Ca 2+ -containing (1.8 mM) media for 30 min and β-hexosaminidase release was measured

    Techniques Used:

    (A) Codeine causes degranulation by HuMC CD34 + -derived cells and LAD2 cells. HuMC and LAD2 cells were stimulated with codeine for 30 min and β-hexosaminidase release was measured ( n = 3). (B) LAD2 cells were stimulated with 2.5 μg/ml codeine
    Figure Legend Snippet: (A) Codeine causes degranulation by HuMC CD34 + -derived cells and LAD2 cells. HuMC and LAD2 cells were stimulated with codeine for 30 min and β-hexosaminidase release was measured ( n = 3). (B) LAD2 cells were stimulated with 2.5 μg/ml codeine

    Techniques Used: Derivative Assay

    3) Product Images from "Hydrogen Sulfide Improves Cardiomyocyte Function in a Cardiac Arrest Model"

    Article Title: Hydrogen Sulfide Improves Cardiomyocyte Function in a Cardiac Arrest Model

    Journal: Annals of Transplantation

    doi: 10.12659/AOT.901410

    Effect of GYY4137 on ATP content and protein synthesis in HL-1 cells. ( A ) Relative ATP content during preconditioning. ATP content is represented as relative content to non-preconditioned cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with the inactive GYY4137. ( B ) Top panel , dot blot assay showing methionine L-azidohomoalanine (AHA) incorporation at 3 hours of preconditioning. Bottom panel , experimental set up for AHA incorporation: Negative control, non-biotinylated AHA; positive control, exponential cell growth in complete Claycomb medium; no analogue, biotinylation of extract without AHA incorporation; actidione, exponential cell growth in complete Claycomb medium after pre-treatment with the protein synthesis inhibitor cycloheximide. ( C ) Relative ATP content during nutrient starvation. ATP content is represented as relative content to the initial level of control cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with inactive GYY4137. ( D ) Dot blot assay showing AHA incorporation at 30 minutes (0.5 hours), 1 hour, and 3 hours after starvation. * p
    Figure Legend Snippet: Effect of GYY4137 on ATP content and protein synthesis in HL-1 cells. ( A ) Relative ATP content during preconditioning. ATP content is represented as relative content to non-preconditioned cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with the inactive GYY4137. ( B ) Top panel , dot blot assay showing methionine L-azidohomoalanine (AHA) incorporation at 3 hours of preconditioning. Bottom panel , experimental set up for AHA incorporation: Negative control, non-biotinylated AHA; positive control, exponential cell growth in complete Claycomb medium; no analogue, biotinylation of extract without AHA incorporation; actidione, exponential cell growth in complete Claycomb medium after pre-treatment with the protein synthesis inhibitor cycloheximide. ( C ) Relative ATP content during nutrient starvation. ATP content is represented as relative content to the initial level of control cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with inactive GYY4137. ( D ) Dot blot assay showing AHA incorporation at 30 minutes (0.5 hours), 1 hour, and 3 hours after starvation. * p

    Techniques Used: Dot Blot, Negative Control, Positive Control

    GYY4137 improves cardioplegic cardio-protective performance during cardiac arrest. ( A ) Scheme of the procedure to test cardioplegic solutions in rat hearts utilizing a Langendorff apparatus. ( B ) Western blotting for caspase-3 in rat heart tissues perfused with Conv or del Nido cardioplegic solutions with or without GYY4137 (n=4). The graph shows quantification of densitometry analysis utilizing ImageJ software. ( C ) ATP quantification in rat heart tissue after finalization of Langendorff perfusion protocol as described in methods section. For the sham group, rat hearts were not subjected to the experimental Langendorff protocol; the organs were directly explanted and ATP content was analyzed. * p
    Figure Legend Snippet: GYY4137 improves cardioplegic cardio-protective performance during cardiac arrest. ( A ) Scheme of the procedure to test cardioplegic solutions in rat hearts utilizing a Langendorff apparatus. ( B ) Western blotting for caspase-3 in rat heart tissues perfused with Conv or del Nido cardioplegic solutions with or without GYY4137 (n=4). The graph shows quantification of densitometry analysis utilizing ImageJ software. ( C ) ATP quantification in rat heart tissue after finalization of Langendorff perfusion protocol as described in methods section. For the sham group, rat hearts were not subjected to the experimental Langendorff protocol; the organs were directly explanted and ATP content was analyzed. * p

    Techniques Used: Western Blot, Software

    Effect of GYY4137 on HL-1 cell apoptosis. ( A ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in glucose and amino acid deprivation medium (-Glu KH). ( B ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in amino acid deprivation medium (KH). ( C, D ) Effect of GYY4137 preconditioning on apoptosis of cells treated with staurosporine ( C ) and doxorubicin ( D ). ( E ) Effect of GYY4137 on apoptosis of cells during starvation in -Glu KH medium without preconditioning. In all cases, results show percentage of apoptosis over time. Dashed lines represent cells treated with active GYY4137 and solid lines represent cells treated with inactive GYY4137. Three replicates in three differences experiments were performed per condition. * p
    Figure Legend Snippet: Effect of GYY4137 on HL-1 cell apoptosis. ( A ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in glucose and amino acid deprivation medium (-Glu KH). ( B ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in amino acid deprivation medium (KH). ( C, D ) Effect of GYY4137 preconditioning on apoptosis of cells treated with staurosporine ( C ) and doxorubicin ( D ). ( E ) Effect of GYY4137 on apoptosis of cells during starvation in -Glu KH medium without preconditioning. In all cases, results show percentage of apoptosis over time. Dashed lines represent cells treated with active GYY4137 and solid lines represent cells treated with inactive GYY4137. Three replicates in three differences experiments were performed per condition. * p

    Techniques Used: Cell Culture

    Short-term electrical recovery after GYY4137 treatment in the isolated Langendorff-perfused rat model. ( A ) Representative 5 second volume-conducted pseudo-ECG traces showing pre-treatment (Pre, light) or post-treatment (Post, dark) with cardioplegic solutions. A better preservation of heart rate (HR) and more regular cardiac activation is detected with del Nido+GYY4137 formulation than with untreated KH solution-preserved hearts. Traces are segmented after 15 minutes of registration from a single-lead electrode located at the epicardial base of the right ventricle in the Langendorff-perfused whole-heart. ( B ) Quantification of the effects of cardioplegic treatment on heart rate (HR, beats per minute) reveals significant slowing of cardiac activation, whereas heartrate-slowing is non-significantly different in the del Nido + GYY4137 treated group. * p
    Figure Legend Snippet: Short-term electrical recovery after GYY4137 treatment in the isolated Langendorff-perfused rat model. ( A ) Representative 5 second volume-conducted pseudo-ECG traces showing pre-treatment (Pre, light) or post-treatment (Post, dark) with cardioplegic solutions. A better preservation of heart rate (HR) and more regular cardiac activation is detected with del Nido+GYY4137 formulation than with untreated KH solution-preserved hearts. Traces are segmented after 15 minutes of registration from a single-lead electrode located at the epicardial base of the right ventricle in the Langendorff-perfused whole-heart. ( B ) Quantification of the effects of cardioplegic treatment on heart rate (HR, beats per minute) reveals significant slowing of cardiac activation, whereas heartrate-slowing is non-significantly different in the del Nido + GYY4137 treated group. * p

    Techniques Used: Isolation, Preserving, Activation Assay

    GYY4137 reduces oxidative stress during cardiac arrest. ( A ) Levels of oxidative stress biomarkers GSH, GSSG; and ( B ) SAM and SAH were analyzed in rat hearts treated with Conv or del Nido cardioplegia solutions with or without GYY4137. Graphs show the ratio of GSH/GSSG ( A ) and SAM/SAH ( B ). Four-six experiments were performed per condition. * p
    Figure Legend Snippet: GYY4137 reduces oxidative stress during cardiac arrest. ( A ) Levels of oxidative stress biomarkers GSH, GSSG; and ( B ) SAM and SAH were analyzed in rat hearts treated with Conv or del Nido cardioplegia solutions with or without GYY4137. Graphs show the ratio of GSH/GSSG ( A ) and SAM/SAH ( B ). Four-six experiments were performed per condition. * p

    Techniques Used:

    4) Product Images from "Circulating mitochondrial DNA and Toll-like receptor 9 are associated with vascular dysfunction in spontaneously hypertensive rats"

    Article Title: Circulating mitochondrial DNA and Toll-like receptor 9 are associated with vascular dysfunction in spontaneously hypertensive rats

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvv137

    Cyclooxygenase (COX) and p38 MAPK inhibition normalizes ODN2395-induced sensitivity to NE in MRA. Concentration-response curves for NE, with or without indomethacin (10 −5 mol/L), in MRA from ( A ) Veh- and ( B ) ODN2395-treated rats. ( C ) Area under
    Figure Legend Snippet: Cyclooxygenase (COX) and p38 MAPK inhibition normalizes ODN2395-induced sensitivity to NE in MRA. Concentration-response curves for NE, with or without indomethacin (10 −5 mol/L), in MRA from ( A ) Veh- and ( B ) ODN2395-treated rats. ( C ) Area under

    Techniques Used: Inhibition, Concentration Assay

    Acute incubation of isolated MRA with TLR9 agonist ODN2395 activates TLR9 signalling. Incubation of naïve Sprague Dawley MRA with TLR9 agonist ODN2395 (2 µmol/L) for 15 or 30 min did not change protein expression of ( A ) TLR9. However,
    Figure Legend Snippet: Acute incubation of isolated MRA with TLR9 agonist ODN2395 activates TLR9 signalling. Incubation of naïve Sprague Dawley MRA with TLR9 agonist ODN2395 (2 µmol/L) for 15 or 30 min did not change protein expression of ( A ) TLR9. However,

    Techniques Used: Incubation, Isolation, Expressing

    TLR9 agonist ODN2395 impairs endothelium-dependent relaxation in MRA and renders MRA more sensitive to NE via decreased nitric oxide and increased reactive oxygen species. Concentration-response curves for ( A ) acetylcholine (ACh), ( B ) sodium nitroprusside
    Figure Legend Snippet: TLR9 agonist ODN2395 impairs endothelium-dependent relaxation in MRA and renders MRA more sensitive to NE via decreased nitric oxide and increased reactive oxygen species. Concentration-response curves for ( A ) acetylcholine (ACh), ( B ) sodium nitroprusside

    Techniques Used: Concentration Assay

    TLR9 antagonist ODN2088 lowers SBP in SHR, and TLR9 agonist ODN2395 increases SBP in previously normotensive rats. Systolic blood pressure responses ( A ) across ODN2088 treatment in WKY and SHR (arrows depict when ODN2088 or Veh were administered) and
    Figure Legend Snippet: TLR9 antagonist ODN2088 lowers SBP in SHR, and TLR9 agonist ODN2395 increases SBP in previously normotensive rats. Systolic blood pressure responses ( A ) across ODN2088 treatment in WKY and SHR (arrows depict when ODN2088 or Veh were administered) and

    Techniques Used:

    5) Product Images from "Mosaic Labeling and 3-Dimensional Morphological Analysis of Single Cells in the Zebrafish Left-right Organizer"

    Article Title: Mosaic Labeling and 3-Dimensional Morphological Analysis of Single Cells in the Zebrafish Left-right Organizer

    Journal: Bio-protocol

    doi: 10.21769/BioProtoc.3090

    Mosaic labeling and 3D rendering of single KV cells. A. Double transgenic Tg(sox17:Cre ERT2 ); Tg(ubi:Zebrabow) zebrafish are incrossed to obtain embryos. B. Time course of mosaic labeling of KV cells. Brief treatment of double transgenic Tg(sox17:Cre ERT2 ); Tg(ubi:Zebrabow) embryos with 4-OHT from the dome stage (4 h post-fertilization) to the shield stage (6 hpf) generates low levels of Cre activity that changes expression of default RFP to expression of CFP or YFP in a subset of KV cells. C. Structure of the ubi:zebrabow and sox17:Cre ERT2 transgenes and the possible recombination outcomes of the Zebrabow transgene by Cre recombinase activity in KV cell lineages. Cre can mediate the deletion of sequences flanked by loxP sites (orange triangles) or variant lox2272 sites (blue triangles), leaving behind single loxP or lox2272 sites that are not cross-compatible with each other. D. Mosaic labeled YFP + KV cells (pseudo-colored green) at the middle plane of KV at tailbud stage and 8 somite stage (8 ss). Scale bars = 20 μm. E. 3D reconstructions of single KV cells (green) using Imaris software at tailbud and 8 ss. Dashed line indicates KV lumen surface. Scale bars = 10 μm.
    Figure Legend Snippet: Mosaic labeling and 3D rendering of single KV cells. A. Double transgenic Tg(sox17:Cre ERT2 ); Tg(ubi:Zebrabow) zebrafish are incrossed to obtain embryos. B. Time course of mosaic labeling of KV cells. Brief treatment of double transgenic Tg(sox17:Cre ERT2 ); Tg(ubi:Zebrabow) embryos with 4-OHT from the dome stage (4 h post-fertilization) to the shield stage (6 hpf) generates low levels of Cre activity that changes expression of default RFP to expression of CFP or YFP in a subset of KV cells. C. Structure of the ubi:zebrabow and sox17:Cre ERT2 transgenes and the possible recombination outcomes of the Zebrabow transgene by Cre recombinase activity in KV cell lineages. Cre can mediate the deletion of sequences flanked by loxP sites (orange triangles) or variant lox2272 sites (blue triangles), leaving behind single loxP or lox2272 sites that are not cross-compatible with each other. D. Mosaic labeled YFP + KV cells (pseudo-colored green) at the middle plane of KV at tailbud stage and 8 somite stage (8 ss). Scale bars = 20 μm. E. 3D reconstructions of single KV cells (green) using Imaris software at tailbud and 8 ss. Dashed line indicates KV lumen surface. Scale bars = 10 μm.

    Techniques Used: Labeling, Transgenic Assay, Activity Assay, Expressing, Variant Assay, Software

    6) Product Images from "Ca2+-dependent Conformational Changes in a C-terminal Cytosolic Domain of Polycystin-2 *"

    Article Title: Ca2+-dependent Conformational Changes in a C-terminal Cytosolic Domain of Polycystin-2 *

    Journal:

    doi: 10.1074/jbc.M109.025635

    Tertiary Structure of Polycystin-2-(680–796)
    Figure Legend Snippet: Tertiary Structure of Polycystin-2-(680–796)

    Techniques Used:

    Expression and Spectroscopic Characterization of the C-terminal Fragments of Human Polycystin-2
    Figure Legend Snippet: Expression and Spectroscopic Characterization of the C-terminal Fragments of Human Polycystin-2

    Techniques Used: Expressing

    C-terminal domain of polycystin-2. Fragment of the C-terminal sequence of polycystin-2-(680–796) investigated in this report. Italics , amino acids from the thrombin cleavage site. An EF-hand is predicted by Prosite, with the central, metal binding
    Figure Legend Snippet: C-terminal domain of polycystin-2. Fragment of the C-terminal sequence of polycystin-2-(680–796) investigated in this report. Italics , amino acids from the thrombin cleavage site. An EF-hand is predicted by Prosite, with the central, metal binding

    Techniques Used: Sequencing, Binding Assay

    Aggregation state of polycystin-2-(680–796). Plot of I / I ref as a function of the relative gradient strength G with I denoting the signal intensity at gradient G and I ref the intensity at the lowest gradient strength. The samples contained 200
    Figure Legend Snippet: Aggregation state of polycystin-2-(680–796). Plot of I / I ref as a function of the relative gradient strength G with I denoting the signal intensity at gradient G and I ref the intensity at the lowest gradient strength. The samples contained 200

    Techniques Used:

    Manganese binding to polycystin-2-(680–796). To a sample of 500 μ m unlabeled polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 100 m m NaCl, 2 m m dithioerythritol, 0.1 m m DSS, 5 m m CaCl 2 in 90% H 2 O, 10% D 2 O a concentrated solution
    Figure Legend Snippet: Manganese binding to polycystin-2-(680–796). To a sample of 500 μ m unlabeled polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 100 m m NaCl, 2 m m dithioerythritol, 0.1 m m DSS, 5 m m CaCl 2 in 90% H 2 O, 10% D 2 O a concentrated solution

    Techniques Used: Binding Assay

    CD spectra of polycystin-2-(680–796). The sample contained 424 μ m polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 500 m m NaCl, before ( thin ) and after ( bold ) the addition of 2 m m CaCl 2 . The data were fitted using the online server
    Figure Legend Snippet: CD spectra of polycystin-2-(680–796). The sample contained 424 μ m polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 500 m m NaCl, before ( thin ) and after ( bold ) the addition of 2 m m CaCl 2 . The data were fitted using the online server

    Techniques Used:

    1 H- 15 N HSQC spectra of polycystin-2-(680–796). The sample conditions were: 0.25 m m polycystin-2-(680–796) in 10 m m potassium phosphate buffer, pH 6.8, 500 m m NaCl, 2 m m dithioerythritol, and 0.1 m m DSS containing 90% H 2 O and 10% D 2 O. Temperature
    Figure Legend Snippet: 1 H- 15 N HSQC spectra of polycystin-2-(680–796). The sample conditions were: 0.25 m m polycystin-2-(680–796) in 10 m m potassium phosphate buffer, pH 6.8, 500 m m NaCl, 2 m m dithioerythritol, and 0.1 m m DSS containing 90% H 2 O and 10% D 2 O. Temperature

    Techniques Used:

    Gel electrophoresis of the C-terminal fragment of polycystin-2. PAGE of polycystin-2-(680–796) before and after removal of the His tag.
    Figure Legend Snippet: Gel electrophoresis of the C-terminal fragment of polycystin-2. PAGE of polycystin-2-(680–796) before and after removal of the His tag.

    Techniques Used: Nucleic Acid Electrophoresis, Polyacrylamide Gel Electrophoresis

    Calcium binding of polycystin-2-(680–796) detected by fluorescence spectroscopy. The sample contained 51.9 μ m polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 500 m m NaCl. The initial calcium concentration of the sample was 8.4
    Figure Legend Snippet: Calcium binding of polycystin-2-(680–796) detected by fluorescence spectroscopy. The sample contained 51.9 μ m polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 500 m m NaCl. The initial calcium concentration of the sample was 8.4

    Techniques Used: Binding Assay, Fluorescence, Spectroscopy, Concentration Assay

    Ca 2+ affinity measurements by NMR spectroscopy. a , Ca 2+ -induced spectral changes observed in a sample of 40 μ m polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 500 m m NaCl, 2 m m dithioerythritol, and 0.1 m m DSS containing 90% H 2 O and 10%
    Figure Legend Snippet: Ca 2+ affinity measurements by NMR spectroscopy. a , Ca 2+ -induced spectral changes observed in a sample of 40 μ m polycystin-2-(680–796) in 5 m m Tris-HCl, pH 6.8, 500 m m NaCl, 2 m m dithioerythritol, and 0.1 m m DSS containing 90% H 2 O and 10%

    Techniques Used: Nuclear Magnetic Resonance, Spectroscopy

    Calcium-binding sites of polycystin-2-(680–796). a , ribbon presentation of the lowest energy conformation of polycystin-2-(680–796) obtained after AUREMOL-ISIC refinement. The side-chain atoms of residues predicted to interact with Ca
    Figure Legend Snippet: Calcium-binding sites of polycystin-2-(680–796). a , ribbon presentation of the lowest energy conformation of polycystin-2-(680–796) obtained after AUREMOL-ISIC refinement. The side-chain atoms of residues predicted to interact with Ca

    Techniques Used: Binding Assay

    Binding of Divalent Ions to Polycystin-2-(680–796)
    Figure Legend Snippet: Binding of Divalent Ions to Polycystin-2-(680–796)

    Techniques Used: Binding Assay

    Tertiary Structure of Polycystin-2-(680–796)
    Figure Legend Snippet: Tertiary Structure of Polycystin-2-(680–796)

    Techniques Used:

    Well folded domain (residues 728–778) of the three-dimensional structure of polycystin-2-(680–796). a , sausage representation of structural ensemble obtained without AUREMOL-ISIC refinement ( dark blue ) and with AUREMOL-ISIC refinement
    Figure Legend Snippet: Well folded domain (residues 728–778) of the three-dimensional structure of polycystin-2-(680–796). a , sausage representation of structural ensemble obtained without AUREMOL-ISIC refinement ( dark blue ) and with AUREMOL-ISIC refinement

    Techniques Used:

    7) Product Images from "Tracking of enzymatic biomass deconstruction by fungal secretomes highlights markers of lignocellulose recalcitrance"

    Article Title: Tracking of enzymatic biomass deconstruction by fungal secretomes highlights markers of lignocellulose recalcitrance

    Journal: Biotechnology for Biofuels

    doi: 10.1186/s13068-019-1417-8

    PCA of the biomass samples FT-IR spectra. A: R0 and R1, B: R2. Wheat straw (WS), blue; miscanthus (MI), red; poplar (PO), green; Ae, A. elegans ; La, L. arvalis ; Tl, T. ljubarskyi ; Tr, T. reesei . Ellipses were drawn manually to highlight clusters
    Figure Legend Snippet: PCA of the biomass samples FT-IR spectra. A: R0 and R1, B: R2. Wheat straw (WS), blue; miscanthus (MI), red; poplar (PO), green; Ae, A. elegans ; La, L. arvalis ; Tl, T. ljubarskyi ; Tr, T. reesei . Ellipses were drawn manually to highlight clusters

    Techniques Used:

    8) Product Images from "Mangiferin Prevents Guinea Pig Tracheal Contraction via Activation of the Nitric Oxide-Cyclic GMP Pathway"

    Article Title: Mangiferin Prevents Guinea Pig Tracheal Contraction via Activation of the Nitric Oxide-Cyclic GMP Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071759

    Proposed mechanism of action of mangiferin on guinea pig tracheal epithelium and smooth muscle cells. Mangiferin activates nitric oxide synthase 3 (NOS3) isoform that up-regulates the production of nitric oxide (NO) in the epithelial cell. Nitric oxide activates guanylate cyclase (GC), which increases the level of intracellular cyclic GMP (cGMP). Increased cGMP then activates the protein kinase G (PKG) cascade, enhancing K + efflux and attenuating Ca 2+ influx-associated smooth muscle cell contractility. GTP (guanosine triphosphate); L-NAME (N-nitro-L-arginine methyl ester); ODQ (1H- [1] , [2] , [4] oxadiazolo[4,3-a]quinoxalin-1-one); TEA (tetraethylammonium).
    Figure Legend Snippet: Proposed mechanism of action of mangiferin on guinea pig tracheal epithelium and smooth muscle cells. Mangiferin activates nitric oxide synthase 3 (NOS3) isoform that up-regulates the production of nitric oxide (NO) in the epithelial cell. Nitric oxide activates guanylate cyclase (GC), which increases the level of intracellular cyclic GMP (cGMP). Increased cGMP then activates the protein kinase G (PKG) cascade, enhancing K + efflux and attenuating Ca 2+ influx-associated smooth muscle cell contractility. GTP (guanosine triphosphate); L-NAME (N-nitro-L-arginine methyl ester); ODQ (1H- [1] , [2] , [4] oxadiazolo[4,3-a]quinoxalin-1-one); TEA (tetraethylammonium).

    Techniques Used:

    9) Product Images from "Targeting microbial biofilms using Ficin, a nonspecific plant protease"

    Article Title: Targeting microbial biofilms using Ficin, a nonspecific plant protease

    Journal: Scientific Reports

    doi: 10.1038/srep46068

    The biofilm formation by S. aureus and S. epidermidis cultivated in Basal medium (BM), Luria-Bertani broth (LB), Müller-Hinton broth (MH), or Trypticase soy broth (TSB) on 35-mm polystyrol adhesive plates. 72 hours-old biofilms were stained by crystal violet.
    Figure Legend Snippet: The biofilm formation by S. aureus and S. epidermidis cultivated in Basal medium (BM), Luria-Bertani broth (LB), Müller-Hinton broth (MH), or Trypticase soy broth (TSB) on 35-mm polystyrol adhesive plates. 72 hours-old biofilms were stained by crystal violet.

    Techniques Used: Staining

    10) Product Images from "Btk-dependent Rac activation and actin rearrangement following Fc?RI aggregation promotes enhanced chemotactic responses of mast cells"

    Article Title: Btk-dependent Rac activation and actin rearrangement following Fc?RI aggregation promotes enhanced chemotactic responses of mast cells

    Journal: Journal of Cell Science

    doi: 10.1242/jcs.071043

    Synergistic cell migration is primarily dependent on chemotaxis. ( A ) IgE-sensitized BMMCs were washed three times with HEPES buffer containing 0.5% BSA, and then cells were stimulated with indicated agonists [antigen, Ag (10 ng/ml), SCF (10 ng/ml), adenosine (1 μM), PGE 2 (100 nM)]. After 3 hours, cell-free supernatants (Sup) from antigen and/or other agonist-stimulated BMMCs were applied to the lower chamber. Sensitized or unsensitized BMMCs were placed in the upper chambers. After incubation for 4 hours, BMMCs migrating to the lower chambers were collected and counted. ( B ) Sensitized BMMCs were preincubated with or without actinomycin D (5 μg/ml) for 30 minutes, and cell migration was measured. ( C ) To test whether the cell migration was chemotaxis or chemokinesis, indicated agonists were placed in the lower chamber or both upper and lower chambers. Sensitized BMMCs were placed in upper chambers. After 4 hours, BMMCs migrating to the lower chambers were collected and counted. Results are means ± s.e. of three separate experiments. * P
    Figure Legend Snippet: Synergistic cell migration is primarily dependent on chemotaxis. ( A ) IgE-sensitized BMMCs were washed three times with HEPES buffer containing 0.5% BSA, and then cells were stimulated with indicated agonists [antigen, Ag (10 ng/ml), SCF (10 ng/ml), adenosine (1 μM), PGE 2 (100 nM)]. After 3 hours, cell-free supernatants (Sup) from antigen and/or other agonist-stimulated BMMCs were applied to the lower chamber. Sensitized or unsensitized BMMCs were placed in the upper chambers. After incubation for 4 hours, BMMCs migrating to the lower chambers were collected and counted. ( B ) Sensitized BMMCs were preincubated with or without actinomycin D (5 μg/ml) for 30 minutes, and cell migration was measured. ( C ) To test whether the cell migration was chemotaxis or chemokinesis, indicated agonists were placed in the lower chamber or both upper and lower chambers. Sensitized BMMCs were placed in upper chambers. After 4 hours, BMMCs migrating to the lower chambers were collected and counted. Results are means ± s.e. of three separate experiments. * P

    Techniques Used: Migration, Chemotaxis Assay, Incubation

    11) Product Images from "Design and Investigation of Optical Properties of N-(Rhodamine-B)-Lactam-Ethylenediamine (RhB-EDA) Fluorescent Probe"

    Article Title: Design and Investigation of Optical Properties of N-(Rhodamine-B)-Lactam-Ethylenediamine (RhB-EDA) Fluorescent Probe

    Journal: Sensors (Basel, Switzerland)

    doi: 10.3390/s18041201

    Fluorescence response of Rhodamine B (10 −7 M) at 575 nm (λ ex = 555 nm) as a function of various added metal cations (10 −3 M) in aqueous solution of 100 mM phosphate buffer (pH 7). From left to right: K + , Mg 2+ , Cu 2+ , Ni 2+ , Fe 2+ , Pb 2+ , Na + , Mn 2+ , Li + , Al 3+ , Co 2+ , Hg 2+ , Sr 2+ , Ca 2+ , Ag + , Cd 2+ , Zn 2+ . λ ex /λ em was selected at 555/575 nm. Slit ex/em 6.0/6.0 nm.
    Figure Legend Snippet: Fluorescence response of Rhodamine B (10 −7 M) at 575 nm (λ ex = 555 nm) as a function of various added metal cations (10 −3 M) in aqueous solution of 100 mM phosphate buffer (pH 7). From left to right: K + , Mg 2+ , Cu 2+ , Ni 2+ , Fe 2+ , Pb 2+ , Na + , Mn 2+ , Li + , Al 3+ , Co 2+ , Hg 2+ , Sr 2+ , Ca 2+ , Ag + , Cd 2+ , Zn 2+ . λ ex /λ em was selected at 555/575 nm. Slit ex/em 6.0/6.0 nm.

    Techniques Used: Fluorescence

    12) Product Images from "Phytosulfokine-? Controls Hypocotyl Length and Cell Expansion in Arabidopsis thaliana through Phytosulfokine Receptor 1"

    Article Title: Phytosulfokine-? Controls Hypocotyl Length and Cell Expansion in Arabidopsis thaliana through Phytosulfokine Receptor 1

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0021054

    Protoplasts from the Arabidopsis hypocotyl expand in response to PSK-α. Protoplasts were isolated from etiolated hypocotyls and their volume was determined at 5 min intervals. After 30 min, at t = 0 min, effectors were added and protoplast volumes were recorded for another 35 min. (A) Addition of 0.1 nM or 1 µM PSK-α caused a rapid and continuous increase in protoplast volume whereas unsulfated PSK peptide (usPSK) or 100 nM of the sulfated peptide CCK8 did not (n = 3–5, P
    Figure Legend Snippet: Protoplasts from the Arabidopsis hypocotyl expand in response to PSK-α. Protoplasts were isolated from etiolated hypocotyls and their volume was determined at 5 min intervals. After 30 min, at t = 0 min, effectors were added and protoplast volumes were recorded for another 35 min. (A) Addition of 0.1 nM or 1 µM PSK-α caused a rapid and continuous increase in protoplast volume whereas unsulfated PSK peptide (usPSK) or 100 nM of the sulfated peptide CCK8 did not (n = 3–5, P

    Techniques Used: Isolation

    13) Product Images from "Laforin, the most common protein mutated in Lafora disease, regulates autophagy"

    Article Title: Laforin, the most common protein mutated in Lafora disease, regulates autophagy

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddq190

    Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.

    Techniques Used: Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.

    Techniques Used: Activity Assay, Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.

    Techniques Used: Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.

    Techniques Used: Activity Assay, Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    14) Product Images from "Distinct PGE2-responder and non-responder phenotypes in human mast cell populations: "All or nothing" enhancement of antigen-dependent mediator release"

    Article Title: Distinct PGE2-responder and non-responder phenotypes in human mast cell populations: "All or nothing" enhancement of antigen-dependent mediator release

    Journal: Immunology letters

    doi: 10.1016/j.imlet.2011.07.002

    Expression of EP receptors in BMMCs, LAD2 cells or HuMCs. A. RNA and cDNA were prepared from the cells as described in ‘Material and Methods’, and real time PCR was performed using commercial Taq-Man EP probes. The expression was normalized
    Figure Legend Snippet: Expression of EP receptors in BMMCs, LAD2 cells or HuMCs. A. RNA and cDNA were prepared from the cells as described in ‘Material and Methods’, and real time PCR was performed using commercial Taq-Man EP probes. The expression was normalized

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    cAMP production in response to PGE 2 in mouse BMMCs (A) (insert magnifies cAMP scale from 0 to 10 pmoles), LAD2 cells (B) and HuMCs (C,D). Briefly, the cells were preincubated with the cAMP phosphodiesterase inhibitor, IBMX (100 µM) for 10 min
    Figure Legend Snippet: cAMP production in response to PGE 2 in mouse BMMCs (A) (insert magnifies cAMP scale from 0 to 10 pmoles), LAD2 cells (B) and HuMCs (C,D). Briefly, the cells were preincubated with the cAMP phosphodiesterase inhibitor, IBMX (100 µM) for 10 min

    Techniques Used:

    Comparative effects of PGE 2 on antigen-induced degranulation in BMMCs (A), LAD2 cells (B) or HuMCs (C and D). BMMCs, LAD2 cells or HuMCs from eight different donors were sensitized overnight and then challenged with PGE 2 (100 nM) or antigen (DNP-HSA for
    Figure Legend Snippet: Comparative effects of PGE 2 on antigen-induced degranulation in BMMCs (A), LAD2 cells (B) or HuMCs (C and D). BMMCs, LAD2 cells or HuMCs from eight different donors were sensitized overnight and then challenged with PGE 2 (100 nM) or antigen (DNP-HSA for

    Techniques Used:

    15) Product Images from "Direct IBE fermentation from mandarin orange wastes by combination of Clostridium cellulovorans and Clostridium beijerinckii"

    Article Title: Direct IBE fermentation from mandarin orange wastes by combination of Clostridium cellulovorans and Clostridium beijerinckii

    Journal: AMB Express

    doi: 10.1186/s13568-018-0728-7

    Total sugar concentration in the culture medium containing removed peel ( a ) and strained lees ( b ) degraded with or without C. cellulovorans . Values are mean ± SE of three independent samples. An asterisk indicates a significant difference (p
    Figure Legend Snippet: Total sugar concentration in the culture medium containing removed peel ( a ) and strained lees ( b ) degraded with or without C. cellulovorans . Values are mean ± SE of three independent samples. An asterisk indicates a significant difference (p

    Techniques Used: Concentration Assay

    a Butanol yield in the culture supernatants with C. beijerinckii from removed peel and strained lees with or without C. cellulovorans . Values are mean ± SE of four independent samples. b Concentration of reducing sugar in the culture supernatant from removed peel and strained lees with or without C. cellulovorans . Closed and hatched bars indicate before addition of C. beijerinckii and after addition of C. beijerinckii , respectively. The cultivation time was for 18 days. Values are mean ± SE of five independent samples
    Figure Legend Snippet: a Butanol yield in the culture supernatants with C. beijerinckii from removed peel and strained lees with or without C. cellulovorans . Values are mean ± SE of four independent samples. b Concentration of reducing sugar in the culture supernatant from removed peel and strained lees with or without C. cellulovorans . Closed and hatched bars indicate before addition of C. beijerinckii and after addition of C. beijerinckii , respectively. The cultivation time was for 18 days. Values are mean ± SE of five independent samples

    Techniques Used: Concentration Assay

    a Residual total sugar ratio in the culture medium with C. cellulovorans , where different concentrations of limonene (v/v), 0% (filled circle), 0.01% (×), 0.02% (open triangle), 0.05% (open square), and 0.1% (open circle), was present in the culture medium. b Total sugar concentration at 61 days cultivation. Values are mean ± SE of three independent samples
    Figure Legend Snippet: a Residual total sugar ratio in the culture medium with C. cellulovorans , where different concentrations of limonene (v/v), 0% (filled circle), 0.01% (×), 0.02% (open triangle), 0.05% (open square), and 0.1% (open circle), was present in the culture medium. b Total sugar concentration at 61 days cultivation. Values are mean ± SE of three independent samples

    Techniques Used: Concentration Assay

    16) Product Images from "Internal Structure of Matrix-Type Multilayer Capsules Templated on Porous Vaterite CaCO3 Crystals as Probed by Staining with a Fluorescence Dye"

    Article Title: Internal Structure of Matrix-Type Multilayer Capsules Templated on Porous Vaterite CaCO3 Crystals as Probed by Staining with a Fluorescence Dye

    Journal: Micromachines

    doi: 10.3390/mi9110547

    ( a ) Absorbance spectra of R6G ( i ) in absence of polymers; ( ii ) in presence of PDAD; ( iii ) in presence of PSS; and ( iv ) after incubation with PSS that was further removed by ultracentrifugation. ( b ) Concentration dependence of 10 mM absorbance of freeR6G at 533 nm, R6G in presence of PDAD and after incubation with PSS that was further removed by ultracentrifugation. Twenty-five millimolar TRIS buffer solution pH 7.4 containing 137 mM NaCl was used as a solvent.
    Figure Legend Snippet: ( a ) Absorbance spectra of R6G ( i ) in absence of polymers; ( ii ) in presence of PDAD; ( iii ) in presence of PSS; and ( iv ) after incubation with PSS that was further removed by ultracentrifugation. ( b ) Concentration dependence of 10 mM absorbance of freeR6G at 533 nm, R6G in presence of PDAD and after incubation with PSS that was further removed by ultracentrifugation. Twenty-five millimolar TRIS buffer solution pH 7.4 containing 137 mM NaCl was used as a solvent.

    Techniques Used: Incubation, Concentration Assay

    17) Product Images from "Mangiferin Prevents Guinea Pig Tracheal Contraction via Activation of the Nitric Oxide-Cyclic GMP Pathway"

    Article Title: Mangiferin Prevents Guinea Pig Tracheal Contraction via Activation of the Nitric Oxide-Cyclic GMP Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071759

    Involvement of K + channels in the antispasmodic effect of mangiferin. Effects of mangiferin (10 µM) on carbachol-contracted guinea pig trachea, performed in the presence or absence of TEA (A), glibenclamide (B) or apamin (C). Each point represents the mean ± S.E.M. of 6 segments. All results are expressed as a percentage of the contractile response induced by 2.5 µM carbachol. * p
    Figure Legend Snippet: Involvement of K + channels in the antispasmodic effect of mangiferin. Effects of mangiferin (10 µM) on carbachol-contracted guinea pig trachea, performed in the presence or absence of TEA (A), glibenclamide (B) or apamin (C). Each point represents the mean ± S.E.M. of 6 segments. All results are expressed as a percentage of the contractile response induced by 2.5 µM carbachol. * p

    Techniques Used:

    18) Product Images from "Intercalation of calcein into layered silicate magadiite and their optical properties"

    Article Title: Intercalation of calcein into layered silicate magadiite and their optical properties

    Journal: Royal Society Open Science

    doi: 10.1098/rsos.171258

    Elemental mapping images of Ca-magadiite–calcein ( a – f ), Zn-magadiite–calcein ( g – l ) and Al-magadiite–calcein ( m – r ).
    Figure Legend Snippet: Elemental mapping images of Ca-magadiite–calcein ( a – f ), Zn-magadiite–calcein ( g – l ) and Al-magadiite–calcein ( m – r ).

    Techniques Used:

    ( a – c ) FTIR spectra of magadiite, H-magadiite, calcein, Ca, Zn, Al ion-exchanged magadiites and their intercalated compounds; ( d ) Comparative FTIR spectra of calcein, Ca-magadiite–calcein, Zn-magadiite–calcein and Al-magadiite–calcein.
    Figure Legend Snippet: ( a – c ) FTIR spectra of magadiite, H-magadiite, calcein, Ca, Zn, Al ion-exchanged magadiites and their intercalated compounds; ( d ) Comparative FTIR spectra of calcein, Ca-magadiite–calcein, Zn-magadiite–calcein and Al-magadiite–calcein.

    Techniques Used:

    ( a – c ) XRD patterns of magadiite, H-magadiite, calcein, Ca, Zn, Al ion-exchanged magadiites and their intercalated compounds.
    Figure Legend Snippet: ( a – c ) XRD patterns of magadiite, H-magadiite, calcein, Ca, Zn, Al ion-exchanged magadiites and their intercalated compounds.

    Techniques Used:

    Inverted fluorescence microscope images of Ca-magadiite–calcein ( a ), Zn-magadiite–calcein ( b ) and Al-magadiite–calcein ( c ) under 250 nm UV light irradiation.
    Figure Legend Snippet: Inverted fluorescence microscope images of Ca-magadiite–calcein ( a ), Zn-magadiite–calcein ( b ) and Al-magadiite–calcein ( c ) under 250 nm UV light irradiation.

    Techniques Used: Fluorescence, Microscopy, Irradiation

    XPS spectra of Ca-magadiite–calcein ( a ), Zn-magadiite–calcein ( b ) and Al-magadiite–calcein ( c ); inserting core-level spectra of Ca 2p , Zn 2p , Al 2p .
    Figure Legend Snippet: XPS spectra of Ca-magadiite–calcein ( a ), Zn-magadiite–calcein ( b ) and Al-magadiite–calcein ( c ); inserting core-level spectra of Ca 2p , Zn 2p , Al 2p .

    Techniques Used:

    A schematic illustration of synthesis of intercalation of calcein into the interlayer spaces of Ca-, Zn-, Al-magadiites (Ca-magadiite–calcein, Zn-magadiite–calcein and Al-magadiite–calcein).
    Figure Legend Snippet: A schematic illustration of synthesis of intercalation of calcein into the interlayer spaces of Ca-, Zn-, Al-magadiites (Ca-magadiite–calcein, Zn-magadiite–calcein and Al-magadiite–calcein).

    Techniques Used:

    Molecular structure of calcein.
    Figure Legend Snippet: Molecular structure of calcein.

    Techniques Used:

    Fluorescence spectra of calcein, Ca-magadiite–calcein, Zn-magadiite–calcein and Al-magadiite–calcein.
    Figure Legend Snippet: Fluorescence spectra of calcein, Ca-magadiite–calcein, Zn-magadiite–calcein and Al-magadiite–calcein.

    Techniques Used: Fluorescence

    TG curves of ( a ) Ca-magadiite and Ca-magadiite–calcein, ( b ) Zn-magadiite and Zn-magadiite–calcein and ( c ) Al-magadiite and Al-magadiite–calcein; inserting their corresponding DTA curves.
    Figure Legend Snippet: TG curves of ( a ) Ca-magadiite and Ca-magadiite–calcein, ( b ) Zn-magadiite and Zn-magadiite–calcein and ( c ) Al-magadiite and Al-magadiite–calcein; inserting their corresponding DTA curves.

    Techniques Used:

    SEM images of Ca-magadiite–calcein ( a , b ), Zn-magadiite–calcein ( c , d ) and Al-magadiite–calcein ( e , f ).
    Figure Legend Snippet: SEM images of Ca-magadiite–calcein ( a , b ), Zn-magadiite–calcein ( c , d ) and Al-magadiite–calcein ( e , f ).

    Techniques Used:

    19) Product Images from "Developmental Toxicity of Diclofenac and Elucidation of Gene Regulation in zebrafish (Danio rerio)"

    Article Title: Developmental Toxicity of Diclofenac and Elucidation of Gene Regulation in zebrafish (Danio rerio)

    Journal: Scientific Reports

    doi: 10.1038/srep04841

    Distribution of diclofenac in the cytoplasm and membrane o f zebrafish embryos after 8 h exposure. The concentration of diclofenac was between 5 and 162 μM. (A) plots of γ vs C 0 ; (B) plots of γ vs C f .
    Figure Legend Snippet: Distribution of diclofenac in the cytoplasm and membrane o f zebrafish embryos after 8 h exposure. The concentration of diclofenac was between 5 and 162 μM. (A) plots of γ vs C 0 ; (B) plots of γ vs C f .

    Techniques Used: Concentration Assay

    Binding number of diclofenac to the zebrafish embryo with C 0 increasing from 0.3–1620 μM. (A) a, partitioning stage; b, Freundlich adsorption stage; (B) Plot of γ vs C f ( C 0 from 0 to 162 μM); (C) Plot of log γ vs log C f ( C 0 from 162 to 1620 μM).
    Figure Legend Snippet: Binding number of diclofenac to the zebrafish embryo with C 0 increasing from 0.3–1620 μM. (A) a, partitioning stage; b, Freundlich adsorption stage; (B) Plot of γ vs C f ( C 0 from 0 to 162 μM); (C) Plot of log γ vs log C f ( C 0 from 162 to 1620 μM).

    Techniques Used: Binding Assay, Adsorption

    Toxic effect of diclofenac on zebrafish during the exposure at 1–4 dpt. (A) Control group; (B) 1.01 μM exposure group; (C) 3.38 μM exposure group; (D) 10.13 μM exposure group; (E) 15.2 μM exposure group. HE: hemagglutination, MD: muscle degeneration, PE: pericardial edema, SBL: short body length, TC: trunk curvature, TM: tail malformation.
    Figure Legend Snippet: Toxic effect of diclofenac on zebrafish during the exposure at 1–4 dpt. (A) Control group; (B) 1.01 μM exposure group; (C) 3.38 μM exposure group; (D) 10.13 μM exposure group; (E) 15.2 μM exposure group. HE: hemagglutination, MD: muscle degeneration, PE: pericardial edema, SBL: short body length, TC: trunk curvature, TM: tail malformation.

    Techniques Used:

    Illustration of transmembrane transport of diclofenac and its subsequent developmental and genetic toxicity. (The gels and blots of Wnt8a were cropped and the full-length blots/gels were presented in Supplementary Figure S9 ).
    Figure Legend Snippet: Illustration of transmembrane transport of diclofenac and its subsequent developmental and genetic toxicity. (The gels and blots of Wnt8a were cropped and the full-length blots/gels were presented in Supplementary Figure S9 ).

    Techniques Used:

    20) Product Images from "RIPK1 protects hepatocytes from death in Fas-induced hepatitis"

    Article Title: RIPK1 protects hepatocytes from death in Fas-induced hepatitis

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-09789-8

    RIPK1 deficiency increases the sensitivity of primary hepatocytes to Fas-agonist stimulation. Primary cultures of hepatocytes issued from Ripk1 fl/fl ( n = 2) or Ripk1 LPC-KO (n = 2) mice were subjected during 16 h to mAb-Jo2 concentrations ranging from 1 to 5 ng/mL in presence of ETA (1 µg/mL) (left panel). Additional primary cultures (n = 3 for each strains) were exposed to a unique dose of mAb-Jo2 (5 ng/mL) also in presence of ETA (1 µg/mL) for statistical analysis (middle panel). In parallel, these primary hepatocyte cultures were subjected during 16 h to mAb-Jo2 at 20 ng/mL in presence of ETA (1 µg/mL) and in absence or presence of the z-VAD-fmk pan-caspase inhibitor (right panel). Cell death was analysed by WST-1 based assay and data are expressed as a percentage of signal obtained in basal survival conditions without Fas-agonist. Error bars corresponds either to internal triplicates for each primary cultures (left panel) or to triplicates of independent primary cultures (middle and right panels).
    Figure Legend Snippet: RIPK1 deficiency increases the sensitivity of primary hepatocytes to Fas-agonist stimulation. Primary cultures of hepatocytes issued from Ripk1 fl/fl ( n = 2) or Ripk1 LPC-KO (n = 2) mice were subjected during 16 h to mAb-Jo2 concentrations ranging from 1 to 5 ng/mL in presence of ETA (1 µg/mL) (left panel). Additional primary cultures (n = 3 for each strains) were exposed to a unique dose of mAb-Jo2 (5 ng/mL) also in presence of ETA (1 µg/mL) for statistical analysis (middle panel). In parallel, these primary hepatocyte cultures were subjected during 16 h to mAb-Jo2 at 20 ng/mL in presence of ETA (1 µg/mL) and in absence or presence of the z-VAD-fmk pan-caspase inhibitor (right panel). Cell death was analysed by WST-1 based assay and data are expressed as a percentage of signal obtained in basal survival conditions without Fas-agonist. Error bars corresponds either to internal triplicates for each primary cultures (left panel) or to triplicates of independent primary cultures (middle and right panels).

    Techniques Used: Mouse Assay, WST-1 Assay

    21) Product Images from "Visualizing Filamentous Actin Using Phalloidin in Chlamydomonas reinhardtii"

    Article Title: Visualizing Filamentous Actin Using Phalloidin in Chlamydomonas reinhardtii

    Journal: Bio-protocol

    doi: 10.21769/BioProtoc.3274

    Phalloidin staining using non-optimal conditions. A. If cells incubate too long in staining solution or PFA has expired, cells will show strong pyrenoid signal (yellow arrow) and non-specific cytoplasmic staining. This type of signal can be easily distinguished from Atto 488 signal as there will be no filamentous perinuclear or apical staining, instead a dim hazy signal found throughout the entire cell body as pictured in A and B above. Scale bars = 5 μm. C. A general diagram of a Chlamydomonas cell highlighting filamentous actin in pink, the pyrenoid in blue, and the chloroplast in green.
    Figure Legend Snippet: Phalloidin staining using non-optimal conditions. A. If cells incubate too long in staining solution or PFA has expired, cells will show strong pyrenoid signal (yellow arrow) and non-specific cytoplasmic staining. This type of signal can be easily distinguished from Atto 488 signal as there will be no filamentous perinuclear or apical staining, instead a dim hazy signal found throughout the entire cell body as pictured in A and B above. Scale bars = 5 μm. C. A general diagram of a Chlamydomonas cell highlighting filamentous actin in pink, the pyrenoid in blue, and the chloroplast in green.

    Techniques Used: Staining, IF-cells

    22) Product Images from "Vitellogenins in the spider Parasteatoda tepidariorum – expression profile and putative hormonal regulation of vitellogenesis"

    Article Title: Vitellogenins in the spider Parasteatoda tepidariorum – expression profile and putative hormonal regulation of vitellogenesis

    Journal: BMC Developmental Biology

    doi: 10.1186/s12861-019-0184-x

    Level of vitellogenin (at transcript and protein levels) in response to 20-hydroxyecdysone administration. The profile of the vitellogenins [mean ± SD] of the P. tepidariorum females that were treated with a Ringer solution and 10, 100 and 200 ng of 20E per individual. Vg results in the midgut glands at the transcript ( a .1) and protein levels ( a .2), the ovaries at the transcript ( b .1) and protein levels ( b .2) and the hemolymph at the protein level ( c ). The dashed line box shows the physiological time of vitellogenesis. Lowercase letters indicate significant differences between experimental groups at the same time point (Tukey’s multiple comparisons test, p ≤ 0.05)
    Figure Legend Snippet: Level of vitellogenin (at transcript and protein levels) in response to 20-hydroxyecdysone administration. The profile of the vitellogenins [mean ± SD] of the P. tepidariorum females that were treated with a Ringer solution and 10, 100 and 200 ng of 20E per individual. Vg results in the midgut glands at the transcript ( a .1) and protein levels ( a .2), the ovaries at the transcript ( b .1) and protein levels ( b .2) and the hemolymph at the protein level ( c ). The dashed line box shows the physiological time of vitellogenesis. Lowercase letters indicate significant differences between experimental groups at the same time point (Tukey’s multiple comparisons test, p ≤ 0.05)

    Techniques Used:

    23) Product Images from "Rho-kinase-dependent F-actin rearrangement is involved in the inhibition of PI3-kinase/Akt during ischemia-reperfusion-induced endothelial cell apoptosis"

    Article Title: Rho-kinase-dependent F-actin rearrangement is involved in the inhibition of PI3-kinase/Akt during ischemia-reperfusion-induced endothelial cell apoptosis

    Journal: Apoptosis

    doi: 10.1007/s10495-007-0173-6

    Y-27632 and cytochalasinD prevent F-actin bundle formation during I/R. Shown is fluorescent imaging of rhodamin-phalloidin stained F-actin structures and DAPI stained nuclei in non-treated, Y-27632-, cytochalasinD-, latrunculinA- and jasplakinolide-treated control, simulated ischemia and simulated I/R cells. The arrow and * indicate F-actin bundles and F-actin clumps, respectively. Control cells : In non-treated cells, most F-actin bundles were seen in the periphery of the cell. Y-27632-treated cells showed a slight decrease in peripheral F-actin bundles. CytochalasinD-treated cells showed only F-actin clumps. Cells treated with latrunculinA or jasplakinolide showed a slight decrease and increase in F-actin bundles, respectively. Ischemic cells : Non-treated cells showed F-actin bundles throughout the whole cell. Cells treated with Y-27632, cytochalasinD or latrunculinA showed only F-actin clumps. Jasplakinolide-treated cells showed F-actin bundles. I/R cells : Non-treated cells showed F-actin bundles throughout the whole cell. In Y-27632-treated cells, no F-actin bundles were seen. CytochalasinD-treated cells showed also no F-actin bundles, only F-actin clumps were visible. LatrunculinA-treated cells showed some F-actin bundles. Jasplakinolide-treated cells showed as many F-actin bundles as non-treated I/R cells
    Figure Legend Snippet: Y-27632 and cytochalasinD prevent F-actin bundle formation during I/R. Shown is fluorescent imaging of rhodamin-phalloidin stained F-actin structures and DAPI stained nuclei in non-treated, Y-27632-, cytochalasinD-, latrunculinA- and jasplakinolide-treated control, simulated ischemia and simulated I/R cells. The arrow and * indicate F-actin bundles and F-actin clumps, respectively. Control cells : In non-treated cells, most F-actin bundles were seen in the periphery of the cell. Y-27632-treated cells showed a slight decrease in peripheral F-actin bundles. CytochalasinD-treated cells showed only F-actin clumps. Cells treated with latrunculinA or jasplakinolide showed a slight decrease and increase in F-actin bundles, respectively. Ischemic cells : Non-treated cells showed F-actin bundles throughout the whole cell. Cells treated with Y-27632, cytochalasinD or latrunculinA showed only F-actin clumps. Jasplakinolide-treated cells showed F-actin bundles. I/R cells : Non-treated cells showed F-actin bundles throughout the whole cell. In Y-27632-treated cells, no F-actin bundles were seen. CytochalasinD-treated cells showed also no F-actin bundles, only F-actin clumps were visible. LatrunculinA-treated cells showed some F-actin bundles. Jasplakinolide-treated cells showed as many F-actin bundles as non-treated I/R cells

    Techniques Used: Imaging, Staining

    Schematic representation of the experimental protocol. Cells were pre-treated for 1 h with Y-27632, cytochalasinD, latrunculinA, jasplakinolide or wortmannin. One hour of simulated ischemia was followed by 1–24 h of simulated reperfusion. No reperfusion for ATP measurement and F-actin staining, 1 h for ATP measurement, F-actin staining and analysis of Akt activity by western blotting and 24 h for quantification of apoptosis. The drugs were also present during the reperfusion phase
    Figure Legend Snippet: Schematic representation of the experimental protocol. Cells were pre-treated for 1 h with Y-27632, cytochalasinD, latrunculinA, jasplakinolide or wortmannin. One hour of simulated ischemia was followed by 1–24 h of simulated reperfusion. No reperfusion for ATP measurement and F-actin staining, 1 h for ATP measurement, F-actin staining and analysis of Akt activity by western blotting and 24 h for quantification of apoptosis. The drugs were also present during the reperfusion phase

    Techniques Used: Staining, Activity Assay, Western Blot

    24) Product Images from "Carboxylated molecules regulate magnesium content of amorphous calcium carbonates during calcification"

    Article Title: Carboxylated molecules regulate magnesium content of amorphous calcium carbonates during calcification

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

    doi: 10.1073/pnas.0906741106

    Mg/Ca ratio in ACC vs. log( K Mg-ligand / K Ca-ligand ) at 0.025 M oxydiacetic, d -tartaric, citric, glutamic, malonic, and aspartic acids. ( A ) Solution Mg/Ca ratio of 2.0. ( B ) Solution Mg/Ca ratio of 4.0. ( C ) Solution Mg/Ca ratio of 5.0 (modern seawater).
    Figure Legend Snippet: Mg/Ca ratio in ACC vs. log( K Mg-ligand / K Ca-ligand ) at 0.025 M oxydiacetic, d -tartaric, citric, glutamic, malonic, and aspartic acids. ( A ) Solution Mg/Ca ratio of 2.0. ( B ) Solution Mg/Ca ratio of 4.0. ( C ) Solution Mg/Ca ratio of 5.0 (modern seawater).

    Techniques Used:

    Mg/Ca ratio in solution vs. Mg/Ca in ACC for the inorganic control experiments ( A ) for aspartic acid at 0.1, 0.05, and 0.025 M, ( B ) for glutamic acid at 0.1, 0.05, and 0.025 M, ( C ) four carboxylic acids at 0.025 M: oxydiacetic, d -tartaric, citric, and
    Figure Legend Snippet: Mg/Ca ratio in solution vs. Mg/Ca in ACC for the inorganic control experiments ( A ) for aspartic acid at 0.1, 0.05, and 0.025 M, ( B ) for glutamic acid at 0.1, 0.05, and 0.025 M, ( C ) four carboxylic acids at 0.025 M: oxydiacetic, d -tartaric, citric, and

    Techniques Used:

    Mg/Ca ratio in the solid vs. log( K Mg-ligand / K Ca-ligand ) for 0.025 M oxydiacetic, d -tartaric, citric, glutamic, malonic, and aspartic acids. Shaded boxes show compositional ranges of high magnesium calcite and dolomite.
    Figure Legend Snippet: Mg/Ca ratio in the solid vs. log( K Mg-ligand / K Ca-ligand ) for 0.025 M oxydiacetic, d -tartaric, citric, glutamic, malonic, and aspartic acids. Shaded boxes show compositional ranges of high magnesium calcite and dolomite.

    Techniques Used:

    25) Product Images from "Assessment of Surfactant Protein A (SP-A) dependent agglutination"

    Article Title: Assessment of Surfactant Protein A (SP-A) dependent agglutination

    Journal: BMC Pulmonary Medicine

    doi: 10.1186/1471-2466-10-59

    SP-A self-agglutination assay . a) The figure shows a scheme of the SP-A self-agglutination-assay. The streptavidin beads were coupled to biotinylated rabbit anti-goat antibodies which bound goat anti-human SP-A antibodies. These anti-SP-A antibodies bound SP-A at its N-terminal end, so SP-A could self-agglutinate by its CRD. The scheme attempts to illustrate the components of the reactants, but does not render how the molecules bind exactly. b) The microscope pictures with a magnification of 10 times were taken under a light microscope. Picture a shows an agglutinate while picture b illustrates beads without agglutination. c) The graph illustrates the SP-A dependency of the bead agglutination. The agglutinate size is plotted against the amount of serum containing SP-A which was incubated with the beads, anti-goat antibody and goat-anti-human antibody in a buffer with calcium ions. Two different sera were used (Black triangle, SP-A concentration 20 ng/ml, sample 2 and balck square, SP-A concentration 21 ng/ml, sample 1) in the experiments which were repeated three times. The final SP-A amount was in 0.6 μl serum about 16 pg.
    Figure Legend Snippet: SP-A self-agglutination assay . a) The figure shows a scheme of the SP-A self-agglutination-assay. The streptavidin beads were coupled to biotinylated rabbit anti-goat antibodies which bound goat anti-human SP-A antibodies. These anti-SP-A antibodies bound SP-A at its N-terminal end, so SP-A could self-agglutinate by its CRD. The scheme attempts to illustrate the components of the reactants, but does not render how the molecules bind exactly. b) The microscope pictures with a magnification of 10 times were taken under a light microscope. Picture a shows an agglutinate while picture b illustrates beads without agglutination. c) The graph illustrates the SP-A dependency of the bead agglutination. The agglutinate size is plotted against the amount of serum containing SP-A which was incubated with the beads, anti-goat antibody and goat-anti-human antibody in a buffer with calcium ions. Two different sera were used (Black triangle, SP-A concentration 20 ng/ml, sample 2 and balck square, SP-A concentration 21 ng/ml, sample 1) in the experiments which were repeated three times. The final SP-A amount was in 0.6 μl serum about 16 pg.

    Techniques Used: Agglutination, Microscopy, Light Microscopy, Incubation, Concentration Assay

    SP-A self-agglutination and fractions . The graphs show the self-agglutination ability (y-axis) of different SP-A structures derived from BAL (b) and serum (a) (x-axis) of the study populations. The streptavidin beads were coupled to biotinylated rabbit anti-goat antibodies which bound goat anti-human SP-A antibodies. These anti-SP-A antibodies bound SP-A at its N-terminal end, so SP-A could self-agglutinate by its CRD. The SP-A amount was adjusted to 1 ng (final concentration 100 ng/ml). All experiments were analyzed by One way ANOVA. * stands for p
    Figure Legend Snippet: SP-A self-agglutination and fractions . The graphs show the self-agglutination ability (y-axis) of different SP-A structures derived from BAL (b) and serum (a) (x-axis) of the study populations. The streptavidin beads were coupled to biotinylated rabbit anti-goat antibodies which bound goat anti-human SP-A antibodies. These anti-SP-A antibodies bound SP-A at its N-terminal end, so SP-A could self-agglutinate by its CRD. The SP-A amount was adjusted to 1 ng (final concentration 100 ng/ml). All experiments were analyzed by One way ANOVA. * stands for p

    Techniques Used: Agglutination, Derivative Assay, Concentration Assay

    26) Product Images from "Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) Reduces Oxidative and Mechanical Stress-Evoked Matrix Degradation in Chondrifying Cell Cultures"

    Article Title: Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) Reduces Oxidative and Mechanical Stress-Evoked Matrix Degradation in Chondrifying Cell Cultures

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20010168

    mRNA ( A ) and protein ( B ) expression of matrix metalloproteinases in chondrifying micromass cultures. Optical densities of signals were measured and results were normalized to the optical densities of 0-day cultures. In panels ( A , B ), the numbers below the signals represent integrated densities of signals determined by ImageJ freeware. For RT-PCR and Western blot reactions, GAPDH ( A ) and actin ( B ) were used as internal controls. Zymography ( C ) with collagen type I, gelatin, and casein substrates was also performed. Signals for MMP9 at 75 kDa, MMP13 at 54 kDa, proMMP9 at 85 kDa, and MMP1 at 54 kDa are labeled by arrows. Densities of three independent experiments are shown in the figures. Asterisks indicate significant differences compared to the 0-day cultures (* p
    Figure Legend Snippet: mRNA ( A ) and protein ( B ) expression of matrix metalloproteinases in chondrifying micromass cultures. Optical densities of signals were measured and results were normalized to the optical densities of 0-day cultures. In panels ( A , B ), the numbers below the signals represent integrated densities of signals determined by ImageJ freeware. For RT-PCR and Western blot reactions, GAPDH ( A ) and actin ( B ) were used as internal controls. Zymography ( C ) with collagen type I, gelatin, and casein substrates was also performed. Signals for MMP9 at 75 kDa, MMP13 at 54 kDa, proMMP9 at 85 kDa, and MMP1 at 54 kDa are labeled by arrows. Densities of three independent experiments are shown in the figures. Asterisks indicate significant differences compared to the 0-day cultures (* p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Zymography, Labeling

    Effects of PACAP and/or oxidative stress and/or mechanical stress (MS) on mRNA ( A ) and protein ( B ) expression of matrix metalloproteinases in chondrifying micromass cultures. Optical densities of signals were measured and results were normalized to the optical densities of three-day cultures ( C ). In panels ( A , B ), the numbers below the signals represent integrated densities of signals determined by ImageJ freeware. For RT-PCR and Western blot reactions, GAPDH ( A ) and actin ( B ) were used as internal controls. Zymography ( C ) with collagen type I, gelatin, and casein substrates was also performed during PACAP treatments, oxidative stress, mechanical stress, and combinations of the three. Signals for MMP9 at 75 kDa, MMP13 at 54 kDa, proMMP9 at 85 kDa, and MMP1 at 54 kDa are labeled. Densities and means of three independent experiments (±standard error of the mean) are shown in the figures. Asterisks indicate significant differences compared with the three-day cultures ( p
    Figure Legend Snippet: Effects of PACAP and/or oxidative stress and/or mechanical stress (MS) on mRNA ( A ) and protein ( B ) expression of matrix metalloproteinases in chondrifying micromass cultures. Optical densities of signals were measured and results were normalized to the optical densities of three-day cultures ( C ). In panels ( A , B ), the numbers below the signals represent integrated densities of signals determined by ImageJ freeware. For RT-PCR and Western blot reactions, GAPDH ( A ) and actin ( B ) were used as internal controls. Zymography ( C ) with collagen type I, gelatin, and casein substrates was also performed during PACAP treatments, oxidative stress, mechanical stress, and combinations of the three. Signals for MMP9 at 75 kDa, MMP13 at 54 kDa, proMMP9 at 85 kDa, and MMP1 at 54 kDa are labeled. Densities and means of three independent experiments (±standard error of the mean) are shown in the figures. Asterisks indicate significant differences compared with the three-day cultures ( p

    Techniques Used: Mass Spectrometry, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Zymography, Labeling

    27) Product Images from "Metabolic pairing of aerobic and anaerobic production in a one-pot batch cultivation"

    Article Title: Metabolic pairing of aerobic and anaerobic production in a one-pot batch cultivation

    Journal: Biotechnology for Biofuels

    doi: 10.1186/s13068-018-1186-9

    TLC analyses demonstrating the wax ester production from ADP1-g— C. butyricum coculture. The wax esters were synthesized by ADP1-g strain utilizing the acetate and butyrate generated during C. butyricum fermentation. The samples are from the same timepoints shown in Fig. 4
    Figure Legend Snippet: TLC analyses demonstrating the wax ester production from ADP1-g— C. butyricum coculture. The wax esters were synthesized by ADP1-g strain utilizing the acetate and butyrate generated during C. butyricum fermentation. The samples are from the same timepoints shown in Fig. 4

    Techniques Used: Thin Layer Chromatography, Synthesized, Generated

    The growth, pH, substrate and metabolite concentrations and hydrogen production from the aerobic–anaerobic phases of the ADP1-g— C. butyricum cocultivations in 300 ml JM medium. Initial aerobic conditions turned to anaerobic via ADP1-g deoxygenation within 12 h of cultivation initiating glucose consumption and hydrogen production by C. butyricum . a OD 600nm (open circle) and pH (closed square) trends of one-pot batch ADP1-g— C. butyricum cocultivations. b Glucose utilization ( C. butyricum, open square), acetate–butyrate metabolism (acetate, open star; butyrate, open triangle) and hydrogen productivity mmol/l/h per culture volume (in bars) from cocultivation experiment. The data points are averaged from triplicate experimental repeats. In some cases, the symbols overlap and the error bars (standard deviation) are smaller than the symbol
    Figure Legend Snippet: The growth, pH, substrate and metabolite concentrations and hydrogen production from the aerobic–anaerobic phases of the ADP1-g— C. butyricum cocultivations in 300 ml JM medium. Initial aerobic conditions turned to anaerobic via ADP1-g deoxygenation within 12 h of cultivation initiating glucose consumption and hydrogen production by C. butyricum . a OD 600nm (open circle) and pH (closed square) trends of one-pot batch ADP1-g— C. butyricum cocultivations. b Glucose utilization ( C. butyricum, open square), acetate–butyrate metabolism (acetate, open star; butyrate, open triangle) and hydrogen productivity mmol/l/h per culture volume (in bars) from cocultivation experiment. The data points are averaged from triplicate experimental repeats. In some cases, the symbols overlap and the error bars (standard deviation) are smaller than the symbol

    Techniques Used: Standard Deviation

    One-pot batch cultivation of ADP1-g and C. butyricum in 1-l bioreactor. The cocultivation was carried out in 800 ml of initially aerobic JM medium supplemented with 20 mM glucose. The changes in ( a ) pH (closed square), pO 2 (asterisk) and biomass formation (in cell dry weight, CDW; open circle), and ( b ) glucose (open square), acetate (open star), butyrate (open triangle) and wax ester concentrations (in bars) are presented. C. butyricum consumed glucose producing acetate and butyrate during the first 24 h (anaerobic phase), whereas ADP1-g utilized the fermentation end-products during the following 10 h (aerobic phase). Arrows indicate the timepoint at which the oxygen supply was initiated and the culture pH adjusted to 7.3. The data for CDW, glucose, acetate and butyrate are mean values from triplicate technical repeats. The standard deviations (error bars) of each data are plotted and in some cases the symbols overlap the error bars
    Figure Legend Snippet: One-pot batch cultivation of ADP1-g and C. butyricum in 1-l bioreactor. The cocultivation was carried out in 800 ml of initially aerobic JM medium supplemented with 20 mM glucose. The changes in ( a ) pH (closed square), pO 2 (asterisk) and biomass formation (in cell dry weight, CDW; open circle), and ( b ) glucose (open square), acetate (open star), butyrate (open triangle) and wax ester concentrations (in bars) are presented. C. butyricum consumed glucose producing acetate and butyrate during the first 24 h (anaerobic phase), whereas ADP1-g utilized the fermentation end-products during the following 10 h (aerobic phase). Arrows indicate the timepoint at which the oxygen supply was initiated and the culture pH adjusted to 7.3. The data for CDW, glucose, acetate and butyrate are mean values from triplicate technical repeats. The standard deviations (error bars) of each data are plotted and in some cases the symbols overlap the error bars

    Techniques Used:

    Schematic representation of the one-pot batch cultivation of C. butyricum and ADP1-g strain. In the first stage, the metabolic activity of ADP1-g deoxidizes the culture, allowing H 2 production from glucose by C. butyricum . Thereafter, oxygen is released to the culture, enabling wax ester production from the residual carbon, namely acetate and butyrate, by ADP1-g
    Figure Legend Snippet: Schematic representation of the one-pot batch cultivation of C. butyricum and ADP1-g strain. In the first stage, the metabolic activity of ADP1-g deoxidizes the culture, allowing H 2 production from glucose by C. butyricum . Thereafter, oxygen is released to the culture, enabling wax ester production from the residual carbon, namely acetate and butyrate, by ADP1-g

    Techniques Used: Activity Assay

    The growth, pH, and substrate concentrations of ADP1-g— C. butyricum cocultivations in 300 ml JM medium. Arrows indicate the time when the growth vessel caps were closed (for anaerobic phase) and opened (for second aerobic phase) during the experiment. a OD 600nm (open circle) and pH (closed square) trends of one-pot batch ADP1-g— C. butyricum cocultivations. b Glucose utilization ( C. butyricum, open square) and acetate–butyrate metabolism (acetate, open star; butyrate, open triangle) data from cocultivation experiment. The data points are averaged from triplicate experimental repeats. In some cases, the symbols overlap and the error bars (standard deviation) are smaller than the symbol
    Figure Legend Snippet: The growth, pH, and substrate concentrations of ADP1-g— C. butyricum cocultivations in 300 ml JM medium. Arrows indicate the time when the growth vessel caps were closed (for anaerobic phase) and opened (for second aerobic phase) during the experiment. a OD 600nm (open circle) and pH (closed square) trends of one-pot batch ADP1-g— C. butyricum cocultivations. b Glucose utilization ( C. butyricum, open square) and acetate–butyrate metabolism (acetate, open star; butyrate, open triangle) data from cocultivation experiment. The data points are averaged from triplicate experimental repeats. In some cases, the symbols overlap and the error bars (standard deviation) are smaller than the symbol

    Techniques Used: Standard Deviation

    28) Product Images from "Characterization of Pediococcus ethanolidurans CUPV141: A β-D-glucan- and Heteropolysaccharide-Producing Bacterium"

    Article Title: Characterization of Pediococcus ethanolidurans CUPV141: A β-D-glucan- and Heteropolysaccharide-Producing Bacterium

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02041

    Adhesion of P. ethanolidurans CUPV141 and CUPV141NR strains to Caco-2 cells. Values are expressed as the percentage of cfu added to the assay. The results are the mean of three independent experiments. The insets show electron micrographs of the bacteria. The arrows mark the β-glucan (EPS). Statistical significances are represented by different letters that mean a p ≤ 0.05.
    Figure Legend Snippet: Adhesion of P. ethanolidurans CUPV141 and CUPV141NR strains to Caco-2 cells. Values are expressed as the percentage of cfu added to the assay. The results are the mean of three independent experiments. The insets show electron micrographs of the bacteria. The arrows mark the β-glucan (EPS). Statistical significances are represented by different letters that mean a p ≤ 0.05.

    Techniques Used:

    Detection of plasmids of P. ethanolidurans CUPV141 and CUPV141NR strains and of P. parvulus 2.6. (A) Detection of the gtf gene by Southern blot hybridization. Left, analysis in a 0.7% agarose gel of plasmids preparations of LAB strains and of E. coli V517. Right, hybridized membrane of samples transferred from the agarose gel. (B) Depicts the calibration curve for plasmid size determination. Symbols: plasmids from E. coli V517 (◊), P. ethanolidurans (♦) and P. parvulus (♦) strains. (C) Analysis in 0.7% agarose gel of gtf PCR amplicons obtained with genomic DNA from CUPV141 and CUPV141NR strains. Smart Ladder, molecular weight standard.
    Figure Legend Snippet: Detection of plasmids of P. ethanolidurans CUPV141 and CUPV141NR strains and of P. parvulus 2.6. (A) Detection of the gtf gene by Southern blot hybridization. Left, analysis in a 0.7% agarose gel of plasmids preparations of LAB strains and of E. coli V517. Right, hybridized membrane of samples transferred from the agarose gel. (B) Depicts the calibration curve for plasmid size determination. Symbols: plasmids from E. coli V517 (◊), P. ethanolidurans (♦) and P. parvulus (♦) strains. (C) Analysis in 0.7% agarose gel of gtf PCR amplicons obtained with genomic DNA from CUPV141 and CUPV141NR strains. Smart Ladder, molecular weight standard.

    Techniques Used: Southern Blot, Hybridization, Agarose Gel Electrophoresis, Plasmid Preparation, Polymerase Chain Reaction, Molecular Weight

    29) Product Images from "The phosphoinositide 3-kinase-dependent activation of Btk is required for optimal eicosanoid production and generation of reactive oxygen species in antigen-stimulated mast cells"

    Article Title: The phosphoinositide 3-kinase-dependent activation of Btk is required for optimal eicosanoid production and generation of reactive oxygen species in antigen-stimulated mast cells

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    The generation of eicoanoids and ROS (A, B) and phosphorylation of cPLA 2 and ERK1/2 (C, D) in antigen-challenged human (A, C) and mouse (B, D) mast cells. IgE-sensitized HuMCs and BMMCs were stimulated with streptavidin (SA, 10 ng/ml) or DNP-HSA (10 ng/ml) respectively for the indicated times, then cell-free supernatants were analyzed for LTC 4 and PGD 2 content (A, B). ROS generation was monitored by DCF fluorescence at 37 °C for up to 30 min at 1 min intervals (A, B). cPLA 2 and ERK1/2 phosphorylation was determined by immunoblot analysis as discussed in Materials and Methods. Experiments not shown revealed that the apparent delay in the ROS production shown in this and subsequent figures was a technical artifact reflecting the time required for the cells to reach optimal temperature (37°) (C, D). Protein levels were normalized to cPLA 2 or ERK then to antigen responses at 10 min to determine the relative intensities presented under each blot. In A and B, results are means ± S.E. of 3 separate experiments performed in duplicate for LTC 4 and PGD 2 generation and kinetic graphs representative of 3 separate experiments for ROS production. In C and D, the blots are representative of three independent experiments.
    Figure Legend Snippet: The generation of eicoanoids and ROS (A, B) and phosphorylation of cPLA 2 and ERK1/2 (C, D) in antigen-challenged human (A, C) and mouse (B, D) mast cells. IgE-sensitized HuMCs and BMMCs were stimulated with streptavidin (SA, 10 ng/ml) or DNP-HSA (10 ng/ml) respectively for the indicated times, then cell-free supernatants were analyzed for LTC 4 and PGD 2 content (A, B). ROS generation was monitored by DCF fluorescence at 37 °C for up to 30 min at 1 min intervals (A, B). cPLA 2 and ERK1/2 phosphorylation was determined by immunoblot analysis as discussed in Materials and Methods. Experiments not shown revealed that the apparent delay in the ROS production shown in this and subsequent figures was a technical artifact reflecting the time required for the cells to reach optimal temperature (37°) (C, D). Protein levels were normalized to cPLA 2 or ERK then to antigen responses at 10 min to determine the relative intensities presented under each blot. In A and B, results are means ± S.E. of 3 separate experiments performed in duplicate for LTC 4 and PGD 2 generation and kinetic graphs representative of 3 separate experiments for ROS production. In C and D, the blots are representative of three independent experiments.

    Techniques Used: Fluorescence

    30) Product Images from "Expression of miRNA-122 Induced by Liver Toxicants in Zebrafish"

    Article Title: Expression of miRNA-122 Induced by Liver Toxicants in Zebrafish

    Journal: BioMed Research International

    doi: 10.1155/2016/1473578

    Histopathology of the adult liver treated with 0.5 μ M tamoxifen for 24 hours. (a) Control untreated zebrafish liver and (b) 0.5 μ M tamoxifen-treated zebrafish liver. Any significant cell death was not detectable in the 0.5 μ M tamoxifen-treated adult liver, but vacuole formation was detected in hepatocytes. H E staining.
    Figure Legend Snippet: Histopathology of the adult liver treated with 0.5 μ M tamoxifen for 24 hours. (a) Control untreated zebrafish liver and (b) 0.5 μ M tamoxifen-treated zebrafish liver. Any significant cell death was not detectable in the 0.5 μ M tamoxifen-treated adult liver, but vacuole formation was detected in hepatocytes. H E staining.

    Techniques Used: Histopathology, Staining

    Quantitative expression levels of miRNA-122 in zebrafish larvae (5 dpf). Expression of miRNA-122 did not change as a result of 5 μ M tamoxifen in zebrafish larvae. Error bars indicate minimum and maximum values of relative quantification.
    Figure Legend Snippet: Quantitative expression levels of miRNA-122 in zebrafish larvae (5 dpf). Expression of miRNA-122 did not change as a result of 5 μ M tamoxifen in zebrafish larvae. Error bars indicate minimum and maximum values of relative quantification.

    Techniques Used: Expressing

    Tissue-specific cell death in the zebrafish larvae treated with tamoxifen (TAM) or metronidazole (Mtz). (a) 0.1% DMSO-treated control (5 dpf) and (b) 1 μ M and (c) 5 μ M TAM-treated zebrafish larvae. (d) 0.1% DMSO-treated control (2 dpf) and (e) 10 mM Mtz-treated zebrafish larvae. Liver-specific cell death was visualized by reduction of transparency in the TAM-treated zebrafish larvae (red arrow), compared to brain-specific cell death in the Mtz-treated larvae (white asterisk). For Mtz experiments, the transgenic zebrafish system, having neuron-specific nitroreductase expression, was used [ 20 ].
    Figure Legend Snippet: Tissue-specific cell death in the zebrafish larvae treated with tamoxifen (TAM) or metronidazole (Mtz). (a) 0.1% DMSO-treated control (5 dpf) and (b) 1 μ M and (c) 5 μ M TAM-treated zebrafish larvae. (d) 0.1% DMSO-treated control (2 dpf) and (e) 10 mM Mtz-treated zebrafish larvae. Liver-specific cell death was visualized by reduction of transparency in the TAM-treated zebrafish larvae (red arrow), compared to brain-specific cell death in the Mtz-treated larvae (white asterisk). For Mtz experiments, the transgenic zebrafish system, having neuron-specific nitroreductase expression, was used [ 20 ].

    Techniques Used: Transgenic Assay, Expressing

    Duration-dependent changes induced by exposure to 5 μ M tamoxifen in zebrafish larvae. (a) Pretreatment at 4 dpf and (b) 2-hour exposure, (c) 4-hour exposure, (d) 8-hour exposure, (e) 12-hour exposure, and (f) 24-hour exposure. After 12 hours, tamoxifen induced cell death in zebrafish larvae liver.
    Figure Legend Snippet: Duration-dependent changes induced by exposure to 5 μ M tamoxifen in zebrafish larvae. (a) Pretreatment at 4 dpf and (b) 2-hour exposure, (c) 4-hour exposure, (d) 8-hour exposure, (e) 12-hour exposure, and (f) 24-hour exposure. After 12 hours, tamoxifen induced cell death in zebrafish larvae liver.

    Techniques Used:

    Quantitative expression levels of miRNA-122 in liver toxicant-treated adult zebrafish. (a) Quantitative expression levels of miRNA-122 in tamoxifen- (or acetaminophen-) treated adult zebrafish livers. miRNA-122 expression was upregulated at 0.5 μ M and 1 μ M and downregulated at 2 μ M tamoxifen. miRNA-122 expression was gradually upregulated at 1 mM and 2 mM and decreased at 4 mM acetaminophen. (b) Tissue-specific expression of miRNA-122. miRNA-122 was expressed in the liver of tamoxifen-treated zebrafish, but not in the brain, heart, and intestine at 0.5 μ M. Error bars indicate minimum and maximum values of relative quantification ( ∗∗ p
    Figure Legend Snippet: Quantitative expression levels of miRNA-122 in liver toxicant-treated adult zebrafish. (a) Quantitative expression levels of miRNA-122 in tamoxifen- (or acetaminophen-) treated adult zebrafish livers. miRNA-122 expression was upregulated at 0.5 μ M and 1 μ M and downregulated at 2 μ M tamoxifen. miRNA-122 expression was gradually upregulated at 1 mM and 2 mM and decreased at 4 mM acetaminophen. (b) Tissue-specific expression of miRNA-122. miRNA-122 was expressed in the liver of tamoxifen-treated zebrafish, but not in the brain, heart, and intestine at 0.5 μ M. Error bars indicate minimum and maximum values of relative quantification ( ∗∗ p

    Techniques Used: Expressing

    31) Product Images from "RIPK1 protects hepatocytes from death in Fas-induced hepatitis"

    Article Title: RIPK1 protects hepatocytes from death in Fas-induced hepatitis

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-09789-8

    Fas-mediated liver injury in Ripk1 LPC-KO mice is independent of TNF-α. ( a ) Levels of serum ALT and AST (n = 4–10) in Ripk1 LPC-KO mice after 3 or 6 h mAb-Jo2 injection with a possible pre-treatment with ETA (ns: non-significant). Each circle or dot represent an individual. ( b ) Pictures of liver tissue sections, stained by H E (upper panels) or analysed by IHC for cleaved caspase-3 (lower panels), issued from Ripk1 LPC-KO mice, 6 h after mAb-Jo2 injection with a possible pre-treatment with ETA. Signal quantification of cleaved caspase-3 (lower right panel).
    Figure Legend Snippet: Fas-mediated liver injury in Ripk1 LPC-KO mice is independent of TNF-α. ( a ) Levels of serum ALT and AST (n = 4–10) in Ripk1 LPC-KO mice after 3 or 6 h mAb-Jo2 injection with a possible pre-treatment with ETA (ns: non-significant). Each circle or dot represent an individual. ( b ) Pictures of liver tissue sections, stained by H E (upper panels) or analysed by IHC for cleaved caspase-3 (lower panels), issued from Ripk1 LPC-KO mice, 6 h after mAb-Jo2 injection with a possible pre-treatment with ETA. Signal quantification of cleaved caspase-3 (lower right panel).

    Techniques Used: Mouse Assay, AST Assay, Injection, Staining, Immunohistochemistry

    RIPK1 deficiency increases the sensitivity of primary hepatocytes to Fas-agonist stimulation. Primary cultures of hepatocytes issued from Ripk1 fl/fl ( n = 2) or Ripk1 LPC-KO (n = 2) mice were subjected during 16 h to mAb-Jo2 concentrations ranging from 1 to 5 ng/mL in presence of ETA (1 µg/mL) (left panel). Additional primary cultures (n = 3 for each strains) were exposed to a unique dose of mAb-Jo2 (5 ng/mL) also in presence of ETA (1 µg/mL) for statistical analysis (middle panel). In parallel, these primary hepatocyte cultures were subjected during 16 h to mAb-Jo2 at 20 ng/mL in presence of ETA (1 µg/mL) and in absence or presence of the z-VAD-fmk pan-caspase inhibitor (right panel). Cell death was analysed by WST-1 based assay and data are expressed as a percentage of signal obtained in basal survival conditions without Fas-agonist. Error bars corresponds either to internal triplicates for each primary cultures (left panel) or to triplicates of independent primary cultures (middle and right panels).
    Figure Legend Snippet: RIPK1 deficiency increases the sensitivity of primary hepatocytes to Fas-agonist stimulation. Primary cultures of hepatocytes issued from Ripk1 fl/fl ( n = 2) or Ripk1 LPC-KO (n = 2) mice were subjected during 16 h to mAb-Jo2 concentrations ranging from 1 to 5 ng/mL in presence of ETA (1 µg/mL) (left panel). Additional primary cultures (n = 3 for each strains) were exposed to a unique dose of mAb-Jo2 (5 ng/mL) also in presence of ETA (1 µg/mL) for statistical analysis (middle panel). In parallel, these primary hepatocyte cultures were subjected during 16 h to mAb-Jo2 at 20 ng/mL in presence of ETA (1 µg/mL) and in absence or presence of the z-VAD-fmk pan-caspase inhibitor (right panel). Cell death was analysed by WST-1 based assay and data are expressed as a percentage of signal obtained in basal survival conditions without Fas-agonist. Error bars corresponds either to internal triplicates for each primary cultures (left panel) or to triplicates of independent primary cultures (middle and right panels).

    Techniques Used: Mouse Assay, WST-1 Assay

    RIPK1 deficiency sensitizes mice to Fas-mediated liver injuries. ( a ) Levels of serum ALT and AST, 3 and 6 h after mAb-Jo2 injection in Ripk1 fl/fl and Ripk1 LPC-KO mice (n = 6–7). ( b ) Pictures of liver tissue sections, stained by H E (upper panels) or analysed by TUNEL (in red) and DAPI (in blue) immunofluorescence (lower panels) issued from Ripk1 fl/fl and Ripk1 LPC-KO mice, 6 h after mAb-Jo2 injection. Yellow arrows show necrotic areas, PV: portal vein. ( c ) Immunostaining of cleaved caspase-3 in the livers of Ripk1 fl/fl and Ripk1 LPC-KO mice, 6 h after mAb-Jo2 injection. ( d ) Mean levels of cleaved caspase-3 (left panel) and of JNK phosphorylation status (right panel) in the livers of Ripk1 fl/fl (n = 5) and Ripk1 LPC-KO (n = 5) mice, collected 6 h after mAb-Jo2 injection (see corresponding Western blots in Supplementary Fig. S1 ). ( e ) Levels of hepatic IL-1β, IL-6 and TNF-α transcripts in Ripk1 fl/fl or Ripk1 LPC-KO mice, 6 h after PBS (n = 3 mice) or mAb-Jo2 injection (n = 6–7 mice). For all graphs, each circle represents an individual.
    Figure Legend Snippet: RIPK1 deficiency sensitizes mice to Fas-mediated liver injuries. ( a ) Levels of serum ALT and AST, 3 and 6 h after mAb-Jo2 injection in Ripk1 fl/fl and Ripk1 LPC-KO mice (n = 6–7). ( b ) Pictures of liver tissue sections, stained by H E (upper panels) or analysed by TUNEL (in red) and DAPI (in blue) immunofluorescence (lower panels) issued from Ripk1 fl/fl and Ripk1 LPC-KO mice, 6 h after mAb-Jo2 injection. Yellow arrows show necrotic areas, PV: portal vein. ( c ) Immunostaining of cleaved caspase-3 in the livers of Ripk1 fl/fl and Ripk1 LPC-KO mice, 6 h after mAb-Jo2 injection. ( d ) Mean levels of cleaved caspase-3 (left panel) and of JNK phosphorylation status (right panel) in the livers of Ripk1 fl/fl (n = 5) and Ripk1 LPC-KO (n = 5) mice, collected 6 h after mAb-Jo2 injection (see corresponding Western blots in Supplementary Fig. S1 ). ( e ) Levels of hepatic IL-1β, IL-6 and TNF-α transcripts in Ripk1 fl/fl or Ripk1 LPC-KO mice, 6 h after PBS (n = 3 mice) or mAb-Jo2 injection (n = 6–7 mice). For all graphs, each circle represents an individual.

    Techniques Used: Mouse Assay, AST Assay, Injection, Staining, TUNEL Assay, Immunofluorescence, Immunostaining, Western Blot

    32) Product Images from "MicroRNA 139-5p coordinates APLNR-CXCR4 crosstalk during vascular maturation"

    Article Title: MicroRNA 139-5p coordinates APLNR-CXCR4 crosstalk during vascular maturation

    Journal: Nature Communications

    doi: 10.1038/ncomms11268

    APLNR is expressed in newly lumenized vessels and its expression is flow-dependent. ( a ) Aplnr expression as determined by in situ hybridization (blue) in P5 retinal vasculature with Isolectin B4 staining (white). The boxed area in the middle image is magnified on the right to show higher magnification of the adjacent panel. Scale bar, 50 μm. n =3 retinas. ( b ) Apln expression as determined by in situ hybridization (blue) in P5 retinal vasculature with Isolectin B4 staining (white). The boxed area is magnified on the right to show higher magnification of the adjacent panel. Scale bar, 100 μm (low magnification) or 50 μm (high magnification). n= 4 retinas. Tip cells are marked by the asterisks. ( c ) Localization of FITC Dextran in the P5 retinal vasculature (green) with Isolectin B4 (red) and Erg123 (endothelial nuclei) staining (blue). The higher magnification images are shown in the right panels. Scale bar, 50 μm. n= 4 retinas. ( d ) APLNR transcript expression in response to shear stress alone or with concurrent KLF2 , KLF4 or combined knockdown via siRNA. ( e ) APLNR transcript expression in response to shear stress alone or with concurrent ERK5 knockdown via siRNA. ( f ) Aplnrb expression in the trunk and tail of zebrafish embryos at 50 hpf with or without nifedipine (2 h treatment). Arrows (DMSO) and arrowheads (nifedipine) demarcate aplnrb expression. ( g ) Aplnrb expression in the trunk and tail of zebrafish embryos at 48 hpf with sih or control morpholino (MO) injection. Asterisks mark absence of aplnrb staining in intersegmental vessels (ISVs). Staining in the posterior cardinal vein (PCV) is also reduced. In situ hybridization with the pan-endothelial marker vascular endothelial cadherin (ve-cadherin) shows normal vascular morphology in all conditions. Arrows mark intersegmental blood vessels. Scale bar, 50 μm. ** P ≤0.01, *** P ≤0.001, t -test. Error bars represent s.e.m.
    Figure Legend Snippet: APLNR is expressed in newly lumenized vessels and its expression is flow-dependent. ( a ) Aplnr expression as determined by in situ hybridization (blue) in P5 retinal vasculature with Isolectin B4 staining (white). The boxed area in the middle image is magnified on the right to show higher magnification of the adjacent panel. Scale bar, 50 μm. n =3 retinas. ( b ) Apln expression as determined by in situ hybridization (blue) in P5 retinal vasculature with Isolectin B4 staining (white). The boxed area is magnified on the right to show higher magnification of the adjacent panel. Scale bar, 100 μm (low magnification) or 50 μm (high magnification). n= 4 retinas. Tip cells are marked by the asterisks. ( c ) Localization of FITC Dextran in the P5 retinal vasculature (green) with Isolectin B4 (red) and Erg123 (endothelial nuclei) staining (blue). The higher magnification images are shown in the right panels. Scale bar, 50 μm. n= 4 retinas. ( d ) APLNR transcript expression in response to shear stress alone or with concurrent KLF2 , KLF4 or combined knockdown via siRNA. ( e ) APLNR transcript expression in response to shear stress alone or with concurrent ERK5 knockdown via siRNA. ( f ) Aplnrb expression in the trunk and tail of zebrafish embryos at 50 hpf with or without nifedipine (2 h treatment). Arrows (DMSO) and arrowheads (nifedipine) demarcate aplnrb expression. ( g ) Aplnrb expression in the trunk and tail of zebrafish embryos at 48 hpf with sih or control morpholino (MO) injection. Asterisks mark absence of aplnrb staining in intersegmental vessels (ISVs). Staining in the posterior cardinal vein (PCV) is also reduced. In situ hybridization with the pan-endothelial marker vascular endothelial cadherin (ve-cadherin) shows normal vascular morphology in all conditions. Arrows mark intersegmental blood vessels. Scale bar, 50 μm. ** P ≤0.01, *** P ≤0.001, t -test. Error bars represent s.e.m.

    Techniques Used: Expressing, Flow Cytometry, In Situ Hybridization, Staining, Injection, Marker

    33) Product Images from "Transcriptional Response of Zebrafish Embryos Exposed to Neurotoxic Compounds Reveals a Muscle Activity Dependent hspb11 Expression"

    Article Title: Transcriptional Response of Zebrafish Embryos Exposed to Neurotoxic Compounds Reveals a Muscle Activity Dependent hspb11 Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0029063

    APM mediated hspb11 induction depends on nAChR activity and increased intracellular calcium levels. (A) hspb11 expression analysis in ache and sop fixe −/− zebrafish mutant embryos at 48 hpf. (B) hspb11 expression in sop fixe null mutants and siblings (+/? = heterozygous and wildtype) after APM (6 µM) exposure (26–50 hpf). (C) 1 µM Thapsigargin (TG) and 2 mM caffeine (CAF) induce hspb11 expression in zebrafish embryos (exposure period 48–50 hpf). Bars represent the relative gene expression as fold change of the respective untreated control as mean ± standard deviation of three independent replicate exposures Control = ctrl. * P
    Figure Legend Snippet: APM mediated hspb11 induction depends on nAChR activity and increased intracellular calcium levels. (A) hspb11 expression analysis in ache and sop fixe −/− zebrafish mutant embryos at 48 hpf. (B) hspb11 expression in sop fixe null mutants and siblings (+/? = heterozygous and wildtype) after APM (6 µM) exposure (26–50 hpf). (C) 1 µM Thapsigargin (TG) and 2 mM caffeine (CAF) induce hspb11 expression in zebrafish embryos (exposure period 48–50 hpf). Bars represent the relative gene expression as fold change of the respective untreated control as mean ± standard deviation of three independent replicate exposures Control = ctrl. * P

    Techniques Used: Activity Assay, Expressing, Mutagenesis, Standard Deviation

    34) Product Images from "Design and Investigation of Optical Properties of N-(Rhodamine-B)-Lactam-Ethylenediamine (RhB-EDA) Fluorescent Probe"

    Article Title: Design and Investigation of Optical Properties of N-(Rhodamine-B)-Lactam-Ethylenediamine (RhB-EDA) Fluorescent Probe

    Journal: Sensors (Basel, Switzerland)

    doi: 10.3390/s18041201

    Fluorescence spectra of 100 µM RHB-EDA probe in the presence of various concentrations of Ag + ions at pH 7 (DMSO:MOPS:H 2 O = 3:14:13) (λ ex /λ em = 500/538 nm).
    Figure Legend Snippet: Fluorescence spectra of 100 µM RHB-EDA probe in the presence of various concentrations of Ag + ions at pH 7 (DMSO:MOPS:H 2 O = 3:14:13) (λ ex /λ em = 500/538 nm).

    Techniques Used: Fluorescence

    35) Product Images from "Dopamine tone regulates D1 receptor trafficking and delivery in striatal neurons in dopamine transporter-deficient mice"

    Article Title: Dopamine tone regulates D1 receptor trafficking and delivery in striatal neurons in dopamine transporter-deficient mice

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

    doi:

    Quantitative analysis of the subcellular distribution of D1R in DAT +/− mice after 6-OHDA injection. ( a ) Measure of D1R immunoreactivity at the plasma membrane and in the cytoplasm: The number of immunoparticles +/− SEM was counted in relation to the plasma membrane length (Mb IP/100 μm) and to the cytoplasmic surface (Cyt IP/100 μm 2 ) in cell bodies and dendrites. Cell bodies in 6-OHDA-injected side display decreased cytoplasmic D1R and increased D1R at the plasma membrane. Dendrites display unchanged membrane-bound receptor but increased cytoplasmic D1R. ( b ) Measure of D1R immunoreactivity in the cytoplasmic organelles in the cell bodies: the endoplasmic reticulum (er) and the vesicles (ves) display a significant decrease of D1R in 6-OHDA-injected side as compared with the control side. **, P ≤ 0.001; *, P ≤ 0.05.
    Figure Legend Snippet: Quantitative analysis of the subcellular distribution of D1R in DAT +/− mice after 6-OHDA injection. ( a ) Measure of D1R immunoreactivity at the plasma membrane and in the cytoplasm: The number of immunoparticles +/− SEM was counted in relation to the plasma membrane length (Mb IP/100 μm) and to the cytoplasmic surface (Cyt IP/100 μm 2 ) in cell bodies and dendrites. Cell bodies in 6-OHDA-injected side display decreased cytoplasmic D1R and increased D1R at the plasma membrane. Dendrites display unchanged membrane-bound receptor but increased cytoplasmic D1R. ( b ) Measure of D1R immunoreactivity in the cytoplasmic organelles in the cell bodies: the endoplasmic reticulum (er) and the vesicles (ves) display a significant decrease of D1R in 6-OHDA-injected side as compared with the control side. **, P ≤ 0.001; *, P ≤ 0.05.

    Techniques Used: Mouse Assay, Injection

    Immunohistochemical detection of D1R at the light and electron microscopic level in heterozygous mice unilaterally treated with 6–0HDA. In the control side ( a and c ), striatal neurons display D1R mostly located in the cytoplasm, associated with the Golgi apparatus (Go), the endoplasmic reticulum (er), and vesicles (arrows). In the 6-OHDA-injected side ( b and d ), D1R appears homogeneously distributed in the neuropile and largely redistributed at the plasma membrane of cell bodies ( b , Inset ; and d , arrowheads) with a limited number of immunoparticles in the cytoplasm. Star points to a dendritic profile. (Magnification bar: a and b = 50 μm; a and b , Insets = 10 μm; c and d = 1 μm).
    Figure Legend Snippet: Immunohistochemical detection of D1R at the light and electron microscopic level in heterozygous mice unilaterally treated with 6–0HDA. In the control side ( a and c ), striatal neurons display D1R mostly located in the cytoplasm, associated with the Golgi apparatus (Go), the endoplasmic reticulum (er), and vesicles (arrows). In the 6-OHDA-injected side ( b and d ), D1R appears homogeneously distributed in the neuropile and largely redistributed at the plasma membrane of cell bodies ( b , Inset ; and d , arrowheads) with a limited number of immunoparticles in the cytoplasm. Star points to a dendritic profile. (Magnification bar: a and b = 50 μm; a and b , Insets = 10 μm; c and d = 1 μm).

    Techniques Used: Immunohistochemistry, Mouse Assay, Injection

    Effect of 6-OHDA injection in the MFB on the extracellular dopamine concentration in the striatum of DAT +/− mice. The extracellular dopamine concentration was monitored every 90 s in the striatum with an electrochemically treated carbon fiber electrode combined with differential pulse voltammetry. Injections of the 6-OHDA solution or of the vehicle were performed after a control period of 15 min. The extracellular dopamine level was expressed in percent of the mean of the 10 absolute peak amplitudes recorded before injections. 6–0HDA provokes a rapid and dramatic decrease of the dopamine content as compared with control experiment with vehicle.
    Figure Legend Snippet: Effect of 6-OHDA injection in the MFB on the extracellular dopamine concentration in the striatum of DAT +/− mice. The extracellular dopamine concentration was monitored every 90 s in the striatum with an electrochemically treated carbon fiber electrode combined with differential pulse voltammetry. Injections of the 6-OHDA solution or of the vehicle were performed after a control period of 15 min. The extracellular dopamine level was expressed in percent of the mean of the 10 absolute peak amplitudes recorded before injections. 6–0HDA provokes a rapid and dramatic decrease of the dopamine content as compared with control experiment with vehicle.

    Techniques Used: Injection, Concentration Assay, Mouse Assay

    36) Product Images from "Heterologous Expression of Xylanase Enzymes in Lipogenic Yeast Yarrowia lipolytica"

    Article Title: Heterologous Expression of Xylanase Enzymes in Lipogenic Yeast Yarrowia lipolytica

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0111443

    Growth of xylanase transformants in liquid medium with birchwood xylan as the carbon source. (a) Empty vector, (b) Yl[ThXynII], (c) Yl[AnXlnD], and (d) Yl[ThXynII] and Yl[AnXlnD].
    Figure Legend Snippet: Growth of xylanase transformants in liquid medium with birchwood xylan as the carbon source. (a) Empty vector, (b) Yl[ThXynII], (c) Yl[AnXlnD], and (d) Yl[ThXynII] and Yl[AnXlnD].

    Techniques Used: Plasmid Preparation

    Growth of xylanase transformants in liquid medium with dyed AZCL-birchwood xylan as the carbon source. (a) Empty vector, (b) Yl[ThXynII], (c) Yl[AnXlnD], and (d) Yl[ThXynII] and Yl[AnXlnD].
    Figure Legend Snippet: Growth of xylanase transformants in liquid medium with dyed AZCL-birchwood xylan as the carbon source. (a) Empty vector, (b) Yl[ThXynII], (c) Yl[AnXlnD], and (d) Yl[ThXynII] and Yl[AnXlnD].

    Techniques Used: Plasmid Preparation

    37) Product Images from "Hydrogen Sulfide Improves Cardiomyocyte Function in a Cardiac Arrest Model"

    Article Title: Hydrogen Sulfide Improves Cardiomyocyte Function in a Cardiac Arrest Model

    Journal: Annals of Transplantation

    doi: 10.12659/AOT.901410

    Effect of GYY4137 on ATP content and protein synthesis in HL-1 cells. ( A ) Relative ATP content during preconditioning. ATP content is represented as relative content to non-preconditioned cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with the inactive GYY4137. ( B ) Top panel , dot blot assay showing methionine L-azidohomoalanine (AHA) incorporation at 3 hours of preconditioning. Bottom panel , experimental set up for AHA incorporation: Negative control, non-biotinylated AHA; positive control, exponential cell growth in complete Claycomb medium; no analogue, biotinylation of extract without AHA incorporation; actidione, exponential cell growth in complete Claycomb medium after pre-treatment with the protein synthesis inhibitor cycloheximide. ( C ) Relative ATP content during nutrient starvation. ATP content is represented as relative content to the initial level of control cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with inactive GYY4137. ( D ) Dot blot assay showing AHA incorporation at 30 minutes (0.5 hours), 1 hour, and 3 hours after starvation. * p
    Figure Legend Snippet: Effect of GYY4137 on ATP content and protein synthesis in HL-1 cells. ( A ) Relative ATP content during preconditioning. ATP content is represented as relative content to non-preconditioned cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with the inactive GYY4137. ( B ) Top panel , dot blot assay showing methionine L-azidohomoalanine (AHA) incorporation at 3 hours of preconditioning. Bottom panel , experimental set up for AHA incorporation: Negative control, non-biotinylated AHA; positive control, exponential cell growth in complete Claycomb medium; no analogue, biotinylation of extract without AHA incorporation; actidione, exponential cell growth in complete Claycomb medium after pre-treatment with the protein synthesis inhibitor cycloheximide. ( C ) Relative ATP content during nutrient starvation. ATP content is represented as relative content to the initial level of control cells (0 hour). Dashed line represents cells treated with active GYY4137 and solid line represents cells treated with inactive GYY4137. ( D ) Dot blot assay showing AHA incorporation at 30 minutes (0.5 hours), 1 hour, and 3 hours after starvation. * p

    Techniques Used: Dot Blot, Negative Control, Positive Control

    GYY4137 improves cardioplegic cardio-protective performance during cardiac arrest. ( A ) Scheme of the procedure to test cardioplegic solutions in rat hearts utilizing a Langendorff apparatus. ( B ) Western blotting for caspase-3 in rat heart tissues perfused with Conv or del Nido cardioplegic solutions with or without GYY4137 (n=4). The graph shows quantification of densitometry analysis utilizing ImageJ software. ( C ) ATP quantification in rat heart tissue after finalization of Langendorff perfusion protocol as described in methods section. For the sham group, rat hearts were not subjected to the experimental Langendorff protocol; the organs were directly explanted and ATP content was analyzed. * p
    Figure Legend Snippet: GYY4137 improves cardioplegic cardio-protective performance during cardiac arrest. ( A ) Scheme of the procedure to test cardioplegic solutions in rat hearts utilizing a Langendorff apparatus. ( B ) Western blotting for caspase-3 in rat heart tissues perfused with Conv or del Nido cardioplegic solutions with or without GYY4137 (n=4). The graph shows quantification of densitometry analysis utilizing ImageJ software. ( C ) ATP quantification in rat heart tissue after finalization of Langendorff perfusion protocol as described in methods section. For the sham group, rat hearts were not subjected to the experimental Langendorff protocol; the organs were directly explanted and ATP content was analyzed. * p

    Techniques Used: Western Blot, Software

    Effect of GYY4137 on HL-1 cell apoptosis. ( A ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in glucose and amino acid deprivation medium (-Glu KH). ( B ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in amino acid deprivation medium (KH). ( C, D ) Effect of GYY4137 preconditioning on apoptosis of cells treated with staurosporine ( C ) and doxorubicin ( D ). ( E ) Effect of GYY4137 on apoptosis of cells during starvation in -Glu KH medium without preconditioning. In all cases, results show percentage of apoptosis over time. Dashed lines represent cells treated with active GYY4137 and solid lines represent cells treated with inactive GYY4137. Three replicates in three differences experiments were performed per condition. * p
    Figure Legend Snippet: Effect of GYY4137 on HL-1 cell apoptosis. ( A ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in glucose and amino acid deprivation medium (-Glu KH). ( B ) Effect of GYY4137 preconditioning on apoptosis of cells cultured in amino acid deprivation medium (KH). ( C, D ) Effect of GYY4137 preconditioning on apoptosis of cells treated with staurosporine ( C ) and doxorubicin ( D ). ( E ) Effect of GYY4137 on apoptosis of cells during starvation in -Glu KH medium without preconditioning. In all cases, results show percentage of apoptosis over time. Dashed lines represent cells treated with active GYY4137 and solid lines represent cells treated with inactive GYY4137. Three replicates in three differences experiments were performed per condition. * p

    Techniques Used: Cell Culture

    Short-term electrical recovery after GYY4137 treatment in the isolated Langendorff-perfused rat model. ( A ) Representative 5 second volume-conducted pseudo-ECG traces showing pre-treatment (Pre, light) or post-treatment (Post, dark) with cardioplegic solutions. A better preservation of heart rate (HR) and more regular cardiac activation is detected with del Nido+GYY4137 formulation than with untreated KH solution-preserved hearts. Traces are segmented after 15 minutes of registration from a single-lead electrode located at the epicardial base of the right ventricle in the Langendorff-perfused whole-heart. ( B ) Quantification of the effects of cardioplegic treatment on heart rate (HR, beats per minute) reveals significant slowing of cardiac activation, whereas heartrate-slowing is non-significantly different in the del Nido + GYY4137 treated group. * p
    Figure Legend Snippet: Short-term electrical recovery after GYY4137 treatment in the isolated Langendorff-perfused rat model. ( A ) Representative 5 second volume-conducted pseudo-ECG traces showing pre-treatment (Pre, light) or post-treatment (Post, dark) with cardioplegic solutions. A better preservation of heart rate (HR) and more regular cardiac activation is detected with del Nido+GYY4137 formulation than with untreated KH solution-preserved hearts. Traces are segmented after 15 minutes of registration from a single-lead electrode located at the epicardial base of the right ventricle in the Langendorff-perfused whole-heart. ( B ) Quantification of the effects of cardioplegic treatment on heart rate (HR, beats per minute) reveals significant slowing of cardiac activation, whereas heartrate-slowing is non-significantly different in the del Nido + GYY4137 treated group. * p

    Techniques Used: Isolation, Preserving, Activation Assay

    GYY4137 reduces oxidative stress during cardiac arrest. ( A ) Levels of oxidative stress biomarkers GSH, GSSG; and ( B ) SAM and SAH were analyzed in rat hearts treated with Conv or del Nido cardioplegia solutions with or without GYY4137. Graphs show the ratio of GSH/GSSG ( A ) and SAM/SAH ( B ). Four-six experiments were performed per condition. * p
    Figure Legend Snippet: GYY4137 reduces oxidative stress during cardiac arrest. ( A ) Levels of oxidative stress biomarkers GSH, GSSG; and ( B ) SAM and SAH were analyzed in rat hearts treated with Conv or del Nido cardioplegia solutions with or without GYY4137. Graphs show the ratio of GSH/GSSG ( A ) and SAM/SAH ( B ). Four-six experiments were performed per condition. * p

    Techniques Used:

    38) Product Images from "Laforin, the most common protein mutated in Lafora disease, regulates autophagy"

    Article Title: Laforin, the most common protein mutated in Lafora disease, regulates autophagy

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddq190

    Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.

    Techniques Used: Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.

    Techniques Used: Activity Assay, Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin induces autophagy and facilitates the clearance of autophagy substrates. ( A ) COS-7 or SK-N-SH cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were treated with or without 400 nM bafilomycin A 1 in the last 4 h of the 24 h post-transfection period. Overexpression of wild-type laforin increased autophagosome synthesis, as analysed by immunoblotting with anti-LC3 antibody (upper gels: low exposure (exp.), lower gels: high exposure) and densitometric analysis of LC3-II levels relative to tubulin. ( B ) COS-7 cells, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of transfected (EGFP-positive) cells with EGFP-LC3 vesicles. ( C ) COS-7 cells, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates and cell death at 48 h post-transfection. Overexpression of wild-type laforin reduced the percentages of tranfected (EGFP-positive) cells with mutant huntingtin aggregates and cell death, assessed by apoptotic nuclear morphology (see Materials and Methods). ( D ) atg5 +/+ and atg5 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in atg5 +/+ MEFs, but not in atg5 −/− MEFs. atg5 −/− MEFs had increased aggregates compared with atg5 +/+ MEFs.

    Techniques Used: Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.
    Figure Legend Snippet: Wild-type laforin reduces mTOR activity to regulate autophagy. ( A ) COS-7 cells, transfected with 2 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were analysed for mTOR activity at 24 h post-transfection by immunoblotting with anti-phospho-S6 kinase (P-S6K, Thr389) and anti-phospho-S6 ribosomal protein (P-S6P, Ser235/236) antibodies. Overexpression of wild-type laforin (detected with anti-myc antibody) reduced phosphorylation of S6K and S6P relative to the total proteins. ( B ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-LC3 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-LC3 vesicles at 24 h post-transfection. Overexpression of wild-type laforin increased the proportion of cells with EGFP-LC3 vesicles in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had a lower proportion of cells with EGFP-LC3 vesicles compared with tsc2 +/+ MEFs. ( C ) tsc2 +/+ and tsc2 −/− MEFs, transfected with 0.5 µg EGFP-HDQ74 and either 1.5 µg pcDNA3.1 (empty vector) or Myc-Laforin for 4 h, were fixed and analysed for EGFP-HDQ74 aggregates at 48 h post-transfection. Overexpression of wild-type laforin reduced mutant huntingtin aggregates in tsc2 +/+ MEFs, but not in tsc2 −/− MEFs. tsc2 −/− MEFs had increased aggregates compared with tsc2 +/+ MEFs.

    Techniques Used: Activity Assay, Transfection, Plasmid Preparation, Over Expression, Mutagenesis

    39) Product Images from "Ricin B Chain Targeted to the Endoplasmic Reticulum of Tobacco Protoplasts Is Degraded by a CDC48- and Vacuole-independent Mechanism *Ricin B Chain Targeted to the Endoplasmic Reticulum of Tobacco Protoplasts Is Degraded by a CDC48- and Vacuole-independent Mechanism * "

    Article Title: Ricin B Chain Targeted to the Endoplasmic Reticulum of Tobacco Protoplasts Is Degraded by a CDC48- and Vacuole-independent Mechanism *Ricin B Chain Targeted to the Endoplasmic Reticulum of Tobacco Protoplasts Is Degraded by a CDC48- and Vacuole-independent Mechanism *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M805222200

    Degradation of ricin B chain does not occur in vacuoles. A, protoplasts expressing RTB or phaseolin were subject to pulse-chase as described under “Experimental Procedures.” Where indicated, protoplasts were preincubated for 1 h with 1 μ m bafilomycin A ( baf A ) before radiolabeling. Subsequent IPs were analyzed by SDS-PAGE and fluorography. The arrowhead indicates the size of full-length phaseolin, and the vertical bar indicates vacuolar-generated fragments of phaseolin. B, protoplasts were transfected with vector alone ( vector ), or plasmids encoding RTB, prepro-RTB, or pre-RTB, where RTB is targeted to the ER via the phaseolin signal peptide ( RTB ), or via the native ricin signal peptide followed by a 9-residue propeptide (prepro-RTB) that is removed in vacuoles, or via the native ricin signal peptide alone (pre-RTB). Following pulse-chase, RTB was immunoprecipitated from separated cell homogenates and analyzed by SDS-PAGE and fluorography. In the schematic, SP represents the phaseolin signal peptide; sp represents the ricin signal peptide, and P represents the N-terminal propeptide of the ricin precursor. Numbers at the margins of gels indicate molecular mass markers in kilodaltons.
    Figure Legend Snippet: Degradation of ricin B chain does not occur in vacuoles. A, protoplasts expressing RTB or phaseolin were subject to pulse-chase as described under “Experimental Procedures.” Where indicated, protoplasts were preincubated for 1 h with 1 μ m bafilomycin A ( baf A ) before radiolabeling. Subsequent IPs were analyzed by SDS-PAGE and fluorography. The arrowhead indicates the size of full-length phaseolin, and the vertical bar indicates vacuolar-generated fragments of phaseolin. B, protoplasts were transfected with vector alone ( vector ), or plasmids encoding RTB, prepro-RTB, or pre-RTB, where RTB is targeted to the ER via the phaseolin signal peptide ( RTB ), or via the native ricin signal peptide followed by a 9-residue propeptide (prepro-RTB) that is removed in vacuoles, or via the native ricin signal peptide alone (pre-RTB). Following pulse-chase, RTB was immunoprecipitated from separated cell homogenates and analyzed by SDS-PAGE and fluorography. In the schematic, SP represents the phaseolin signal peptide; sp represents the ricin signal peptide, and P represents the N-terminal propeptide of the ricin precursor. Numbers at the margins of gels indicate molecular mass markers in kilodaltons.

    Techniques Used: Expressing, Pulse Chase, Radioactivity, SDS Page, Generated, Transfection, Plasmid Preparation, Immunoprecipitation

    40) Product Images from "Aldh2 Attenuates Stem Cell Factor/Kit-Dependent Signaling and Activation in Mast Cells"

    Article Title: Aldh2 Attenuates Stem Cell Factor/Kit-Dependent Signaling and Activation in Mast Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20246216

    Aldh2 deficiency enhances mast cell responses to FcεRI and Kit co-stimulation, but not to FcεRI alone. ( A – C ) β-Hexosaminidase release (degranulation) ( A ), and release of the cytokines TNFα ( B ) and IL-6 ( C ), into the media in response to FcεRI stimulation. Aldh2 +/+ and Aldh2 −/− BMMCs were sensitized with anti-Dinitrophenyl (DNP) IgE (100 ng/mL) overnight, washed, and then challenged with the indicated concentrations of Ag (DNP-HSA) in ( A ) or with 25 ng/mL of antigen (Ag) in ( B , C ). ( D ) Degranulation induced by the indicated concentrations of thapsigargin in Aldh2 +/+ and Aldh2 −/− BMMCs. ( E – G ) Degranulation ( E ), and the release of TNFα ( F ) and IL-6 ( G ), induced by the co-stimulation of FcεRI and Kit in Aldh2 +/+ and Aldh2 −/− BMMCs. Sensitized BMMCs were challenged with the indicated concentrations of Ag ( E ) or with 25 ng/mL of Ag in ( F , G ) in the presence of 100 ng/mL SCF. ( H ) IL-6 released by Aldh2 +/+ and Aldh2 −/− BMMCs stimulated only with SCF at the indicated concentrations. Degranulation in ( A , D , E ) is expressed as the percentage of β-hexosaminidase released into the media compared to the total cellular content. Data are the mean ± SEM of five independent cultures. ** p
    Figure Legend Snippet: Aldh2 deficiency enhances mast cell responses to FcεRI and Kit co-stimulation, but not to FcεRI alone. ( A – C ) β-Hexosaminidase release (degranulation) ( A ), and release of the cytokines TNFα ( B ) and IL-6 ( C ), into the media in response to FcεRI stimulation. Aldh2 +/+ and Aldh2 −/− BMMCs were sensitized with anti-Dinitrophenyl (DNP) IgE (100 ng/mL) overnight, washed, and then challenged with the indicated concentrations of Ag (DNP-HSA) in ( A ) or with 25 ng/mL of antigen (Ag) in ( B , C ). ( D ) Degranulation induced by the indicated concentrations of thapsigargin in Aldh2 +/+ and Aldh2 −/− BMMCs. ( E – G ) Degranulation ( E ), and the release of TNFα ( F ) and IL-6 ( G ), induced by the co-stimulation of FcεRI and Kit in Aldh2 +/+ and Aldh2 −/− BMMCs. Sensitized BMMCs were challenged with the indicated concentrations of Ag ( E ) or with 25 ng/mL of Ag in ( F , G ) in the presence of 100 ng/mL SCF. ( H ) IL-6 released by Aldh2 +/+ and Aldh2 −/− BMMCs stimulated only with SCF at the indicated concentrations. Degranulation in ( A , D , E ) is expressed as the percentage of β-hexosaminidase released into the media compared to the total cellular content. Data are the mean ± SEM of five independent cultures. ** p

    Techniques Used:

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    Millipore calcium chloride dihydrate
    Calcium Chloride Dihydrate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore cytokine homogenate buffer
    Leptin and <t>cytokine</t> levels (IL-6 and TNF- α ) after 4 and 6 weeks on the diets. Consistent with the weight gain and epididymal fat pad weights, leptin levels were significantly higher after 4 and 6 weeks in the HAGE-HF group compared to the other groups. Also consistent with the liver histology and presence of inflammation after 6 weeks on the diet, IL-6 and TNF- α levels were significantly higher in the HAGE-HF group after 6 weeks on the diet.
    Cytokine Homogenate Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore cacl2 • 2h2 o
    Leptin and <t>cytokine</t> levels (IL-6 and TNF- α ) after 4 and 6 weeks on the diets. Consistent with the weight gain and epididymal fat pad weights, leptin levels were significantly higher after 4 and 6 weeks in the HAGE-HF group compared to the other groups. Also consistent with the liver histology and presence of inflammation after 6 weeks on the diet, IL-6 and TNF- α levels were significantly higher in the HAGE-HF group after 6 weeks on the diet.
    Cacl2 • 2h2 O, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leptin and cytokine levels (IL-6 and TNF- α ) after 4 and 6 weeks on the diets. Consistent with the weight gain and epididymal fat pad weights, leptin levels were significantly higher after 4 and 6 weeks in the HAGE-HF group compared to the other groups. Also consistent with the liver histology and presence of inflammation after 6 weeks on the diet, IL-6 and TNF- α levels were significantly higher in the HAGE-HF group after 6 weeks on the diet.

    Journal: BioMed Research International

    Article Title: Advanced Glycation End Products Induce Obesity and Hepatosteatosis in CD-1 Wild-Type Mice

    doi: 10.1155/2016/7867852

    Figure Lengend Snippet: Leptin and cytokine levels (IL-6 and TNF- α ) after 4 and 6 weeks on the diets. Consistent with the weight gain and epididymal fat pad weights, leptin levels were significantly higher after 4 and 6 weeks in the HAGE-HF group compared to the other groups. Also consistent with the liver histology and presence of inflammation after 6 weeks on the diet, IL-6 and TNF- α levels were significantly higher in the HAGE-HF group after 6 weeks on the diet.

    Article Snippet: The remaining liver was washed with cold HBSS and suspended in cytokine homogenate buffer (150 mM NaCl, 15 mM Tris, 1 mM CaCl2 ·2H2 O, and 1 mM MgCl2 ·6H2 O, adjusted to pH 7.4) plus 100x protease inhibitor cocktail 1 (Calbiochem, La Jolla, California) to a total volume of 10 mL and homogenized on ice using a Polytron® Homogenizer (Kinematica Inc., Bohemia, NY).

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