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  • 99
    Thermo Fisher mm l l alanyl l glutamine
    Mm L L Alanyl L Glutamine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore l alanyl l glutamine
    L Alanyl L Glutamine, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore l alanyl l alanine lala
    L Alanyl L Alanine Lala, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore l glycyl l histidyl l lysine
    L Glycyl L Histidyl L Lysine, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore phytohemagglutinin l pha l
    Effects of VAEI-treated DC on T cells. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with <t>phytohemagglutinin-L</t> (PC; 10 μg/ml). Unstimulated T cells were further co-cultivated with immature DC (DC, CD4 + ), DC treated with a maturation cocktail (DC Stim, CD4 + ; 500 ng/mL LPS; 50 ng/mL TNF-alpha and 10 ng/mL IL-1beta), VAEI-treated DC (DC + VAEI, CD4 + ; Iscador® Qu Spez; 0.5 μg/ml), DC incubated with ML-depleted VAEI (DC + VAEI-ML, CD4 + ; Iscador®; 0.66 μg/ml) or VAEI and anti-ML antibody (DC + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell proliferation analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data of 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or untreated T cells co-cultured with immature DC (DC, CD4 + ). (C) IFN-γ release by T cells was analyzed in the supernatants of the co-cultures of 6 independent experiments using a cytokine bead array assay. Asterisks indicate significant differences between the groups (*P
    Phytohemagglutinin L Pha L, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Tocris l nitroarginine l nna
    Sources of endogenous agonists acting to increase cGMP in oligodendrocytes in cerebellum. (A) Section from 10-day-old rat cerebellar slice stained for eNOS (red), nNOS (green), and nuclei (DAPI, blue). (B) Section of a slice from a 14-day-old eNOS −/− mouse incubated with BAY 41-2272 (3 μM; 5 min) and stained for CNPase (red) and cGMP (green), showing cGMP accumulation in oligodendrocytes (examples arrowed). Asterisk indicates non-specific staining of blood vessel (staining persisted after omission of primary antibody). (C) cGMP radioimmunoassay measurements in slices of wild-type (WT) and eNOS −/− mouse cerebellum showing responses to BAY41-2272 (red bars) in the absence and presence of <t>L-nitroarginine</t> <t>(L-NNA,</t> 100 μM) and of a mixture of antagonists (“antags,” blue bars) of ionotropic glutamate receptors (100 μM AP5 plus 100 μM CNQX) and of Na + channels (1 μM TTX). Data are from 6–9 slices in three separate experiments; the population means were not significantly different from each other ( P > 0.05 by one-way ANOVA). (D) Cerebellar slices from 10-day-old rats exposed for 5 min to BAY41-2272 (BAY 41; 3 μM, left columns) or YC-1 (50 μM, right columns) under the indicated conditions. The concentrations of AP5, CNQX, TTX and L-NNA were as given above; ODQ was at 10 μM and the general metabotropic glutamate receptor blocker LY341495 was at 100 μM; n = 4-12 (2–5 separate experiments). (E, F) Sections of 10-day-old (E) and 6-week-old (F) rat cerebellar slices exposed to BAY 41-2272 (3 μM, 5 min) in the presence of a mixture of AP5, CNQX and TTX (“antags”), immunostained for cGMP (green) and CNPase (red). (G, H) Immunostained sections of 10-day-old rat cerebellar slices exposed to BAY 41-2272 (5 μM, 5 min) in the presence (G) or absence (H) of Ca 2+ . (I) cGMP levels in the absence (open columns) and presence (red columns) of BAY 41-2272 (3 μM; 5 min) under control and Ca 2+ -free conditions (as indicated); n = 7–9 (three separate experiments). IBMX (1 mM) was present throughout. (J) Images from a 10-day-old rat cerebellar slice immunostained for ANP and/or eNOS as indicated; nuclei are stained with TO-PRO (blue). In the micrographs, arrows point to oligodendrocytes and arrowheads signify putative astrocytes. Key: egl, external granule cell layer; igl, internal granule cell layer; gcl, granule cell layer; ml, molecular layer; Bg, Bergmann glia; wm, white matter. Scale bar (in A) = 100 μm (A); 35 μm (B): 26 μm (E); 50 μm (F, J) and 80 μm (G, H). All images are representative of four slices in two separate experiments. Histogram results (C, D, I) are means ± SEM. [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]
    L Nitroarginine L Nna, supplied by Tocris, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Tocris l ap5
    Diagram representing distributions of the penetrations by segment of the D2 barrel for cells used for analysis in subsequent figures for d <t>-AP5</t> ( left ) and l -AP5 animals ( right ). The penetrations in each group of animals were quite evenly spread through
    L Ap5, supplied by Tocris, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore l canavanine
    Analysis of base editing patterns and efficiencies in single yeast colonies selected for <t>canavanine</t> resistance. A comparison of base editing frequencies for nCDA1-BE3, cCDA1-BE3, and selected base editors with truncated CDA1 domains is shown. Yeast cells were transformed with plasmids expressing the base editor and an sgRNA targeting the Can1–5 site. The target sequence is shown with the cytidines that can potentially undergo editing in red and the PAM in blue. If C-to-T conversion occurs at position −18 or −19 or both, the Can1 gene will be inactivated and the cell becomes resistant to canavanine. Values and error bars reflect the mean and standard deviation of three biological replicates. Source data are provided as a Source Data file. See also Table 1
    L Canavanine, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore viscozyme l
    Analysis of base editing patterns and efficiencies in single yeast colonies selected for <t>canavanine</t> resistance. A comparison of base editing frequencies for nCDA1-BE3, cCDA1-BE3, and selected base editors with truncated CDA1 domains is shown. Yeast cells were transformed with plasmids expressing the base editor and an sgRNA targeting the Can1–5 site. The target sequence is shown with the cytidines that can potentially undergo editing in red and the PAM in blue. If C-to-T conversion occurs at position −18 or −19 or both, the Can1 gene will be inactivated and the cell becomes resistant to canavanine. Values and error bars reflect the mean and standard deviation of three biological replicates. Source data are provided as a Source Data file. See also Table 1
    Viscozyme L, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore l glyceraldehyde
    Analysis of base editing patterns and efficiencies in single yeast colonies selected for <t>canavanine</t> resistance. A comparison of base editing frequencies for nCDA1-BE3, cCDA1-BE3, and selected base editors with truncated CDA1 domains is shown. Yeast cells were transformed with plasmids expressing the base editor and an sgRNA targeting the Can1–5 site. The target sequence is shown with the cytidines that can potentially undergo editing in red and the PAM in blue. If C-to-T conversion occurs at position −18 or −19 or both, the Can1 gene will be inactivated and the cell becomes resistant to canavanine. Values and error bars reflect the mean and standard deviation of three biological replicates. Source data are provided as a Source Data file. See also Table 1
    L Glyceraldehyde, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    TaKaRa bgl l
    Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, <t>Bcl</t> I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.
    Bgl L, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Millipore l rhamnulose
    Gel retardation analysis to evaluate the inhibitory effects of <t>rhamnulose-1-phophate</t> (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR
    L Rhamnulose, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore l fucitol
    Gel retardation analysis to evaluate the inhibitory effects of <t>rhamnulose-1-phophate</t> (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR
    L Fucitol, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Millipore l dops
    Gel retardation analysis to evaluate the inhibitory effects of <t>rhamnulose-1-phophate</t> (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR
    L Dops, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore l ethionine
    Gel retardation analysis to evaluate the inhibitory effects of <t>rhamnulose-1-phophate</t> (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR
    L Ethionine, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore 2mmol l l carnitine
    Gel retardation analysis to evaluate the inhibitory effects of <t>rhamnulose-1-phophate</t> (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR
    2mmol L L Carnitine, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore l kynurenine l kyn
    Treatment with <t>L-Kynurenine</t> fails to inhibit in vivo T CD8 responses to T Ag. WT mice were treated with L-Kyn (30 mg total as described in Materials and Methods) or PBS, and injected i.p. with C57SV cells. Nine days later, splenic T CD8 were examined for IFN-γ accumulation following restimulation with C57SV cells or synthetic peptides corresponding to T Ag epitopes. T Ag-specific T CD8 frequencies were determined after subtracting background and expressed as mean ± SEM of 6 L-Kyn-treated and 8 vehicle-treated mice (A). These values were used to calculate the absolute number of T Ag-specific T CD8 present within each spleen (B).
    L Kynurenine L Kyn, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of VAEI-treated DC on T cells. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Unstimulated T cells were further co-cultivated with immature DC (DC, CD4 + ), DC treated with a maturation cocktail (DC Stim, CD4 + ; 500 ng/mL LPS; 50 ng/mL TNF-alpha and 10 ng/mL IL-1beta), VAEI-treated DC (DC + VAEI, CD4 + ; Iscador® Qu Spez; 0.5 μg/ml), DC incubated with ML-depleted VAEI (DC + VAEI-ML, CD4 + ; Iscador®; 0.66 μg/ml) or VAEI and anti-ML antibody (DC + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell proliferation analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data of 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or untreated T cells co-cultured with immature DC (DC, CD4 + ). (C) IFN-γ release by T cells was analyzed in the supernatants of the co-cultures of 6 independent experiments using a cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Journal: PLoS ONE

    Article Title: Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model

    doi: 10.1371/journal.pone.0181553

    Figure Lengend Snippet: Effects of VAEI-treated DC on T cells. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Unstimulated T cells were further co-cultivated with immature DC (DC, CD4 + ), DC treated with a maturation cocktail (DC Stim, CD4 + ; 500 ng/mL LPS; 50 ng/mL TNF-alpha and 10 ng/mL IL-1beta), VAEI-treated DC (DC + VAEI, CD4 + ; Iscador® Qu Spez; 0.5 μg/ml), DC incubated with ML-depleted VAEI (DC + VAEI-ML, CD4 + ; Iscador®; 0.66 μg/ml) or VAEI and anti-ML antibody (DC + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell proliferation analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data of 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or untreated T cells co-cultured with immature DC (DC, CD4 + ). (C) IFN-γ release by T cells was analyzed in the supernatants of the co-cultures of 6 independent experiments using a cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Article Snippet: As controls, CFSE+ CD4+ T cells were cultured with medium alone (NC) or in the presence of phytohemagglutinin-L (PC; PHA-L; 10 μg/mL; Sigma-Aldrich, Taufkirchen, Germany).

    Techniques: Purification, Cell Culture, Incubation, Staining, Flow Cytometry, Cytometry, Marker, Expressing, Activation Assay

    Effects of VAEI-treated DC on T cells after tumor-induced immunosuppression. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Co-cultivation of T cells was carried out with unstimulated DC treated with 10% RPMI 1640 medium (CD4 + + DC + Medium), 10% tumor supernatant of an RCC line (CD4 + + DC + TS + ), TS and VAEI (CD4 + + DC + TS + VAEI; Iscador® Qu Spez; 0.5 μg/ml), TS and ML-depleted VAEI (CD4 + + DC + TS + VAEI-ML; Iscador®; 0.66 μg/ml) or TS, VAEI and anti-ML antibody (CD4 + + DC + TS + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell division analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data from 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or T cells co-cultured with TS-treated DC (DC + TS). (C) IFN-γ release by T cells was analyzed in the supernatants of 6 independent co-culture experiments using cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Journal: PLoS ONE

    Article Title: Viscum album neutralizes tumor-induced immunosuppression in a human in vitro cell model

    doi: 10.1371/journal.pone.0181553

    Figure Lengend Snippet: Effects of VAEI-treated DC on T cells after tumor-induced immunosuppression. Purified human CD4 + T lymphocytes were cultured in the presence of medium (NC) or stimulated with phytohemagglutinin-L (PC; 10 μg/ml). Co-cultivation of T cells was carried out with unstimulated DC treated with 10% RPMI 1640 medium (CD4 + + DC + Medium), 10% tumor supernatant of an RCC line (CD4 + + DC + TS + ), TS and VAEI (CD4 + + DC + TS + VAEI; Iscador® Qu Spez; 0.5 μg/ml), TS and ML-depleted VAEI (CD4 + + DC + TS + VAEI-ML; Iscador®; 0.66 μg/ml) or TS, VAEI and anti-ML antibody (CD4 + + DC + TS + VAEI + aML-Ab, CD4 + ; 2.5 μg/ml). (A) Cell division analysis was done using CFSE staining and flow cytometry. (B) CD25 surface marker expression was analyzed as a second indicator for T cell activation. Data from 6 individual experiments are presented as mean ± SD in relation to untreated T cells (NC) or T cells co-cultured with TS-treated DC (DC + TS). (C) IFN-γ release by T cells was analyzed in the supernatants of 6 independent co-culture experiments using cytokine bead array assay. Asterisks indicate significant differences between the groups (*P

    Article Snippet: As controls, CFSE+ CD4+ T cells were cultured with medium alone (NC) or in the presence of phytohemagglutinin-L (PC; PHA-L; 10 μg/mL; Sigma-Aldrich, Taufkirchen, Germany).

    Techniques: Purification, Cell Culture, Staining, Flow Cytometry, Cytometry, Marker, Expressing, Activation Assay, Co-Culture Assay

    Sources of endogenous agonists acting to increase cGMP in oligodendrocytes in cerebellum. (A) Section from 10-day-old rat cerebellar slice stained for eNOS (red), nNOS (green), and nuclei (DAPI, blue). (B) Section of a slice from a 14-day-old eNOS −/− mouse incubated with BAY 41-2272 (3 μM; 5 min) and stained for CNPase (red) and cGMP (green), showing cGMP accumulation in oligodendrocytes (examples arrowed). Asterisk indicates non-specific staining of blood vessel (staining persisted after omission of primary antibody). (C) cGMP radioimmunoassay measurements in slices of wild-type (WT) and eNOS −/− mouse cerebellum showing responses to BAY41-2272 (red bars) in the absence and presence of L-nitroarginine (L-NNA, 100 μM) and of a mixture of antagonists (“antags,” blue bars) of ionotropic glutamate receptors (100 μM AP5 plus 100 μM CNQX) and of Na + channels (1 μM TTX). Data are from 6–9 slices in three separate experiments; the population means were not significantly different from each other ( P > 0.05 by one-way ANOVA). (D) Cerebellar slices from 10-day-old rats exposed for 5 min to BAY41-2272 (BAY 41; 3 μM, left columns) or YC-1 (50 μM, right columns) under the indicated conditions. The concentrations of AP5, CNQX, TTX and L-NNA were as given above; ODQ was at 10 μM and the general metabotropic glutamate receptor blocker LY341495 was at 100 μM; n = 4-12 (2–5 separate experiments). (E, F) Sections of 10-day-old (E) and 6-week-old (F) rat cerebellar slices exposed to BAY 41-2272 (3 μM, 5 min) in the presence of a mixture of AP5, CNQX and TTX (“antags”), immunostained for cGMP (green) and CNPase (red). (G, H) Immunostained sections of 10-day-old rat cerebellar slices exposed to BAY 41-2272 (5 μM, 5 min) in the presence (G) or absence (H) of Ca 2+ . (I) cGMP levels in the absence (open columns) and presence (red columns) of BAY 41-2272 (3 μM; 5 min) under control and Ca 2+ -free conditions (as indicated); n = 7–9 (three separate experiments). IBMX (1 mM) was present throughout. (J) Images from a 10-day-old rat cerebellar slice immunostained for ANP and/or eNOS as indicated; nuclei are stained with TO-PRO (blue). In the micrographs, arrows point to oligodendrocytes and arrowheads signify putative astrocytes. Key: egl, external granule cell layer; igl, internal granule cell layer; gcl, granule cell layer; ml, molecular layer; Bg, Bergmann glia; wm, white matter. Scale bar (in A) = 100 μm (A); 35 μm (B): 26 μm (E); 50 μm (F, J) and 80 μm (G, H). All images are representative of four slices in two separate experiments. Histogram results (C, D, I) are means ± SEM. [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Journal: Glia

    Article Title: Nitric oxide targets oligodendrocytes and promotes their morphological differentiation

    doi: 10.1002/glia.22759

    Figure Lengend Snippet: Sources of endogenous agonists acting to increase cGMP in oligodendrocytes in cerebellum. (A) Section from 10-day-old rat cerebellar slice stained for eNOS (red), nNOS (green), and nuclei (DAPI, blue). (B) Section of a slice from a 14-day-old eNOS −/− mouse incubated with BAY 41-2272 (3 μM; 5 min) and stained for CNPase (red) and cGMP (green), showing cGMP accumulation in oligodendrocytes (examples arrowed). Asterisk indicates non-specific staining of blood vessel (staining persisted after omission of primary antibody). (C) cGMP radioimmunoassay measurements in slices of wild-type (WT) and eNOS −/− mouse cerebellum showing responses to BAY41-2272 (red bars) in the absence and presence of L-nitroarginine (L-NNA, 100 μM) and of a mixture of antagonists (“antags,” blue bars) of ionotropic glutamate receptors (100 μM AP5 plus 100 μM CNQX) and of Na + channels (1 μM TTX). Data are from 6–9 slices in three separate experiments; the population means were not significantly different from each other ( P > 0.05 by one-way ANOVA). (D) Cerebellar slices from 10-day-old rats exposed for 5 min to BAY41-2272 (BAY 41; 3 μM, left columns) or YC-1 (50 μM, right columns) under the indicated conditions. The concentrations of AP5, CNQX, TTX and L-NNA were as given above; ODQ was at 10 μM and the general metabotropic glutamate receptor blocker LY341495 was at 100 μM; n = 4-12 (2–5 separate experiments). (E, F) Sections of 10-day-old (E) and 6-week-old (F) rat cerebellar slices exposed to BAY 41-2272 (3 μM, 5 min) in the presence of a mixture of AP5, CNQX and TTX (“antags”), immunostained for cGMP (green) and CNPase (red). (G, H) Immunostained sections of 10-day-old rat cerebellar slices exposed to BAY 41-2272 (5 μM, 5 min) in the presence (G) or absence (H) of Ca 2+ . (I) cGMP levels in the absence (open columns) and presence (red columns) of BAY 41-2272 (3 μM; 5 min) under control and Ca 2+ -free conditions (as indicated); n = 7–9 (three separate experiments). IBMX (1 mM) was present throughout. (J) Images from a 10-day-old rat cerebellar slice immunostained for ANP and/or eNOS as indicated; nuclei are stained with TO-PRO (blue). In the micrographs, arrows point to oligodendrocytes and arrowheads signify putative astrocytes. Key: egl, external granule cell layer; igl, internal granule cell layer; gcl, granule cell layer; ml, molecular layer; Bg, Bergmann glia; wm, white matter. Scale bar (in A) = 100 μm (A); 35 μm (B): 26 μm (E); 50 μm (F, J) and 80 μm (G, H). All images are representative of four slices in two separate experiments. Histogram results (C, D, I) are means ± SEM. [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Article Snippet: ANP, C-type natriuretic peptide (CNP), 8-Bromo-cGMP (8-Br-cGMP), bicuculline, and isobutylmethylxanthine (IBMX) were from Sigma-Aldrich (Dorset, UK); 1H -[1,2,4], oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), L-nitroarginine (L-NNA)), N-methyl-D-aspartate (NMDA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-amino-5-phosphonopentanoic acid (AP5), and 2-[(1S ,2S )-2-carboxycyclopropyl]-3-(9H -xanthen-9-yl)-D-alanine (LY341495) were from Tocris Cookson (Bristol, UK); tetrodotoxin (TTX) was from Latoxan Laboratories (Rosans, France); Vectastain elite ABC KIT was from Vecto Labs Ltd (Peterborough, UK); 3,3′-diaminobenzidine (DAB) was from Sigma-Aldrich (Dorset, UK); Mayer's haemalum was from Raymond A Lamb Ltd (Eastbourne UK).

    Techniques: Staining, Incubation, RIA Assay, Aqueous Normal-phase Chromatography

    Differentiation of oligodendrocytes in cortical cultures and their responsiveness to exogenous and endogenous NO. (A–F) Cultures at 3 DIV (A, B), 6 DIV (C–E), and 17 DIV (F) immunostained for the indicated markers. Arrows point to examples of CNPase-positive oligodendrocytes (A), nNOS-positive cells (B, E), CNPase-positive and/or MBP-positive oligodendrocytes (C), MOG-positive oligodendrocytes (D), and NG2-positive cells (F); arrowheads, examples of GFAP-positive astrocytes (D, F). Scale bar (in A) = 40 μm (for A, C–F) and 29 μm (B). (G–K) cGMP immunostaining (green) in oligodendrocyes (red) immunolabeled with CNPase or MOG at 17 DIV following exposure to PAPA/NO (30 μM, 2 min; G), DEA/NO (100 nM, 2 min; H, I) and/or BAY 41-2272 (1 μM, 7 min; I–K) in the absence (J) or presence (K) of L-nitroarginine (L-NNA, 100 μM), all in the presence of IBMX (1 mM). Arrows, cGMP-positive (G–J) or negative (K) oligodendrocytes; arrowheads, examples of cGMP-positive putative astrocytes (I, J). Scale bar (G) = 40 μm (for G–K). (L) Numbers of pixels containing both red (CNPase) and green (cGMP) signals derived from z-stacks (1 μm) of 0.17 mm 2 images of cultures under the indicated conditions (concentrations and exposure periods as above); data are means ± SEM from three coverslips in two experiments. [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Journal: Glia

    Article Title: Nitric oxide targets oligodendrocytes and promotes their morphological differentiation

    doi: 10.1002/glia.22759

    Figure Lengend Snippet: Differentiation of oligodendrocytes in cortical cultures and their responsiveness to exogenous and endogenous NO. (A–F) Cultures at 3 DIV (A, B), 6 DIV (C–E), and 17 DIV (F) immunostained for the indicated markers. Arrows point to examples of CNPase-positive oligodendrocytes (A), nNOS-positive cells (B, E), CNPase-positive and/or MBP-positive oligodendrocytes (C), MOG-positive oligodendrocytes (D), and NG2-positive cells (F); arrowheads, examples of GFAP-positive astrocytes (D, F). Scale bar (in A) = 40 μm (for A, C–F) and 29 μm (B). (G–K) cGMP immunostaining (green) in oligodendrocyes (red) immunolabeled with CNPase or MOG at 17 DIV following exposure to PAPA/NO (30 μM, 2 min; G), DEA/NO (100 nM, 2 min; H, I) and/or BAY 41-2272 (1 μM, 7 min; I–K) in the absence (J) or presence (K) of L-nitroarginine (L-NNA, 100 μM), all in the presence of IBMX (1 mM). Arrows, cGMP-positive (G–J) or negative (K) oligodendrocytes; arrowheads, examples of cGMP-positive putative astrocytes (I, J). Scale bar (G) = 40 μm (for G–K). (L) Numbers of pixels containing both red (CNPase) and green (cGMP) signals derived from z-stacks (1 μm) of 0.17 mm 2 images of cultures under the indicated conditions (concentrations and exposure periods as above); data are means ± SEM from three coverslips in two experiments. [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Article Snippet: ANP, C-type natriuretic peptide (CNP), 8-Bromo-cGMP (8-Br-cGMP), bicuculline, and isobutylmethylxanthine (IBMX) were from Sigma-Aldrich (Dorset, UK); 1H -[1,2,4], oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), L-nitroarginine (L-NNA)), N-methyl-D-aspartate (NMDA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-amino-5-phosphonopentanoic acid (AP5), and 2-[(1S ,2S )-2-carboxycyclopropyl]-3-(9H -xanthen-9-yl)-D-alanine (LY341495) were from Tocris Cookson (Bristol, UK); tetrodotoxin (TTX) was from Latoxan Laboratories (Rosans, France); Vectastain elite ABC KIT was from Vecto Labs Ltd (Peterborough, UK); 3,3′-diaminobenzidine (DAB) was from Sigma-Aldrich (Dorset, UK); Mayer's haemalum was from Raymond A Lamb Ltd (Eastbourne UK).

    Techniques: Immunostaining, Immunolabeling, Derivative Assay

    Targeting of oligodendrocytes in slices of adult rat cerebellum by exogenous and endogenous NO. Slices were stimulated by (A) PAPA/NO (30 μM, 5 min), (B, C) BAY 41-2272 (3 μM, 5 min), (D) ANP (1 μM, 10 min) or (E) NMDA (30 μM, 2 min) all in the presence of IBMX (1 mM). L-nitroarginine (L-NNA, 100 μM) was present in C and D. Sections were stained for cGMP (green), CNPase (red) and DAPI (blue). Images are all overlays (red/green co-staining appears yellow) and are representative of six slices per condition in two experiments. Key: ml, molecular layer; P, Purkinje cell; Bg, Bergmann glia; gcl, granule cell layer, wm = white matter; arrows, oligodendrocytes; arrowheads, putative astrocytes. Scale bar (A) = 50 μm (applies to all panels). [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Journal: Glia

    Article Title: Nitric oxide targets oligodendrocytes and promotes their morphological differentiation

    doi: 10.1002/glia.22759

    Figure Lengend Snippet: Targeting of oligodendrocytes in slices of adult rat cerebellum by exogenous and endogenous NO. Slices were stimulated by (A) PAPA/NO (30 μM, 5 min), (B, C) BAY 41-2272 (3 μM, 5 min), (D) ANP (1 μM, 10 min) or (E) NMDA (30 μM, 2 min) all in the presence of IBMX (1 mM). L-nitroarginine (L-NNA, 100 μM) was present in C and D. Sections were stained for cGMP (green), CNPase (red) and DAPI (blue). Images are all overlays (red/green co-staining appears yellow) and are representative of six slices per condition in two experiments. Key: ml, molecular layer; P, Purkinje cell; Bg, Bergmann glia; gcl, granule cell layer, wm = white matter; arrows, oligodendrocytes; arrowheads, putative astrocytes. Scale bar (A) = 50 μm (applies to all panels). [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Article Snippet: ANP, C-type natriuretic peptide (CNP), 8-Bromo-cGMP (8-Br-cGMP), bicuculline, and isobutylmethylxanthine (IBMX) were from Sigma-Aldrich (Dorset, UK); 1H -[1,2,4], oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), L-nitroarginine (L-NNA)), N-methyl-D-aspartate (NMDA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-amino-5-phosphonopentanoic acid (AP5), and 2-[(1S ,2S )-2-carboxycyclopropyl]-3-(9H -xanthen-9-yl)-D-alanine (LY341495) were from Tocris Cookson (Bristol, UK); tetrodotoxin (TTX) was from Latoxan Laboratories (Rosans, France); Vectastain elite ABC KIT was from Vecto Labs Ltd (Peterborough, UK); 3,3′-diaminobenzidine (DAB) was from Sigma-Aldrich (Dorset, UK); Mayer's haemalum was from Raymond A Lamb Ltd (Eastbourne UK).

    Techniques: Aqueous Normal-phase Chromatography, Staining

    Endogenous NO targets oligodendrocytes in 8- to 9-day-old rat cerebellar slices. The slices were exposed for 5 min to BAY 41-2272 (3 μM, 5 min; A–D) or YC-1 (50 μM, 5 min; E, F) in the absence (A) or presence (B–F) of IBMX (1 mM). Cryostat sections were immunostained for cGMP (green) together with CNPase (red; A–C, E, F) or GFAP (red; D). Images are all overlays in which green/red co-staining appears yellow. Nuclei are stained with DAPI (blue). The cGMP immunostaining was abolished by L-nitroarginine (L-NNA, 100 μM; C, F). Key: arrows, oligodendrocytes; arrowheads, GFAP-positive astrocytes (D) and putative astrocytes (A, B, E); wm, white matter; igl, internal granule cell layer; P, Purkinje cell; Bg, Bergmann glia. Images are representative of 4–6 slices from two experiments. Scale bar (A) = 25 μm (applies to all panels). [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Journal: Glia

    Article Title: Nitric oxide targets oligodendrocytes and promotes their morphological differentiation

    doi: 10.1002/glia.22759

    Figure Lengend Snippet: Endogenous NO targets oligodendrocytes in 8- to 9-day-old rat cerebellar slices. The slices were exposed for 5 min to BAY 41-2272 (3 μM, 5 min; A–D) or YC-1 (50 μM, 5 min; E, F) in the absence (A) or presence (B–F) of IBMX (1 mM). Cryostat sections were immunostained for cGMP (green) together with CNPase (red; A–C, E, F) or GFAP (red; D). Images are all overlays in which green/red co-staining appears yellow. Nuclei are stained with DAPI (blue). The cGMP immunostaining was abolished by L-nitroarginine (L-NNA, 100 μM; C, F). Key: arrows, oligodendrocytes; arrowheads, GFAP-positive astrocytes (D) and putative astrocytes (A, B, E); wm, white matter; igl, internal granule cell layer; P, Purkinje cell; Bg, Bergmann glia. Images are representative of 4–6 slices from two experiments. Scale bar (A) = 25 μm (applies to all panels). [Color figure can be viewed in the online issue, which is available at http://wileyonlinelibrary.com .]

    Article Snippet: ANP, C-type natriuretic peptide (CNP), 8-Bromo-cGMP (8-Br-cGMP), bicuculline, and isobutylmethylxanthine (IBMX) were from Sigma-Aldrich (Dorset, UK); 1H -[1,2,4], oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), L-nitroarginine (L-NNA)), N-methyl-D-aspartate (NMDA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), 2-amino-5-phosphonopentanoic acid (AP5), and 2-[(1S ,2S )-2-carboxycyclopropyl]-3-(9H -xanthen-9-yl)-D-alanine (LY341495) were from Tocris Cookson (Bristol, UK); tetrodotoxin (TTX) was from Latoxan Laboratories (Rosans, France); Vectastain elite ABC KIT was from Vecto Labs Ltd (Peterborough, UK); 3,3′-diaminobenzidine (DAB) was from Sigma-Aldrich (Dorset, UK); Mayer's haemalum was from Raymond A Lamb Ltd (Eastbourne UK).

    Techniques: Staining, Immunostaining

    Diagram representing distributions of the penetrations by segment of the D2 barrel for cells used for analysis in subsequent figures for d -AP5 ( left ) and l -AP5 animals ( right ). The penetrations in each group of animals were quite evenly spread through

    Journal: The Journal of Neuroscience

    Article Title: Experience-Dependent Plasticity of Adult Rat S1 Cortex Requires Local NMDA Receptor Activation

    doi: 10.1523/JNEUROSCI.18-23-10196.1998

    Figure Lengend Snippet: Diagram representing distributions of the penetrations by segment of the D2 barrel for cells used for analysis in subsequent figures for d -AP5 ( left ) and l -AP5 animals ( right ). The penetrations in each group of animals were quite evenly spread through

    Article Snippet: Both d -AP5 and l -AP5 were purchased from Tocris.

    Techniques:

    Suppression of whisker-driven responses of cells in layers I–IV after perfusion of barrel cortex with d -AP5 at concentrations of 0.1–10 m m . Epidural perfusion was maintained for a minimum of 6 hr. Findings are from 12 animals tested at

    Journal: The Journal of Neuroscience

    Article Title: Experience-Dependent Plasticity of Adult Rat S1 Cortex Requires Local NMDA Receptor Activation

    doi: 10.1523/JNEUROSCI.18-23-10196.1998

    Figure Lengend Snippet: Suppression of whisker-driven responses of cells in layers I–IV after perfusion of barrel cortex with d -AP5 at concentrations of 0.1–10 m m . Epidural perfusion was maintained for a minimum of 6 hr. Findings are from 12 animals tested at

    Article Snippet: Both d -AP5 and l -AP5 were purchased from Tocris.

    Techniques: Whisker Assay

    Top , Distribution of spike durations recorded from d -AP5-perfused animals for the cells in layers II–III ( black bars ) and IV ( gray bars ) of the D2 barrel column used for this study. Note the bimodal nature of the histogram with antimode at 0.75

    Journal: The Journal of Neuroscience

    Article Title: Experience-Dependent Plasticity of Adult Rat S1 Cortex Requires Local NMDA Receptor Activation

    doi: 10.1523/JNEUROSCI.18-23-10196.1998

    Figure Lengend Snippet: Top , Distribution of spike durations recorded from d -AP5-perfused animals for the cells in layers II–III ( black bars ) and IV ( gray bars ) of the D2 barrel column used for this study. Note the bimodal nature of the histogram with antimode at 0.75

    Article Snippet: Both d -AP5 and l -AP5 were purchased from Tocris.

    Techniques:

    Suppression of long-latency responses by perfusion of 500 μ m d -AP5 onto cortex. Cells A and B are two neurons analyzed from barrel D4. Cell A shows changes in mean response magnitudes to center whisker D4 and to surround whiskers D3 and D5 during

    Journal: The Journal of Neuroscience

    Article Title: Experience-Dependent Plasticity of Adult Rat S1 Cortex Requires Local NMDA Receptor Activation

    doi: 10.1523/JNEUROSCI.18-23-10196.1998

    Figure Lengend Snippet: Suppression of long-latency responses by perfusion of 500 μ m d -AP5 onto cortex. Cells A and B are two neurons analyzed from barrel D4. Cell A shows changes in mean response magnitudes to center whisker D4 and to surround whiskers D3 and D5 during

    Article Snippet: Both d -AP5 and l -AP5 were purchased from Tocris.

    Techniques: Whisker Assay

    Analysis of base editing patterns and efficiencies in single yeast colonies selected for canavanine resistance. A comparison of base editing frequencies for nCDA1-BE3, cCDA1-BE3, and selected base editors with truncated CDA1 domains is shown. Yeast cells were transformed with plasmids expressing the base editor and an sgRNA targeting the Can1–5 site. The target sequence is shown with the cytidines that can potentially undergo editing in red and the PAM in blue. If C-to-T conversion occurs at position −18 or −19 or both, the Can1 gene will be inactivated and the cell becomes resistant to canavanine. Values and error bars reflect the mean and standard deviation of three biological replicates. Source data are provided as a Source Data file. See also Table 1

    Journal: Nature Communications

    Article Title: Engineering of high-precision base editors for site-specific single nucleotide replacement

    doi: 10.1038/s41467-018-08034-8

    Figure Lengend Snippet: Analysis of base editing patterns and efficiencies in single yeast colonies selected for canavanine resistance. A comparison of base editing frequencies for nCDA1-BE3, cCDA1-BE3, and selected base editors with truncated CDA1 domains is shown. Yeast cells were transformed with plasmids expressing the base editor and an sgRNA targeting the Can1–5 site. The target sequence is shown with the cytidines that can potentially undergo editing in red and the PAM in blue. If C-to-T conversion occurs at position −18 or −19 or both, the Can1 gene will be inactivated and the cell becomes resistant to canavanine. Values and error bars reflect the mean and standard deviation of three biological replicates. Source data are provided as a Source Data file. See also Table 1

    Article Snippet: The cells were incubated for 20 h prior to plating on YPAD rich or SC media plates without arginine but with 60 mg/mL l -canavanine (Sigma).

    Techniques: Transformation Assay, Expressing, Sequencing, Standard Deviation

    Rigid linkers narrow the width of the editing window of BE3. a Protospacers and PAM (blue) sequences of the genomic loci tested, with the target Cs shown in red. Subscript numbers indicate the positions of the cytidines relative to the PAM. C-to-T editing at any of the indicated Cs inactivates the Can1 transporter and thus causes resistance to canavanine 17 . b Editing efficiency and specificity of the base editors tested as determined by canavanine selection. The x -axis represents the target Cs within the protospacers. The y -axis shows their C-to-T editing frequency (see Methods and Supplementary Figure 1 ). Sequences of canavanine-resistant mutants aligned with the corresponding reference sequences are shown in Supplementary Figure 3 . Values and error bars represent the mean and standard deviation of three independent biological replicates. Source data are provided as a Source Data file

    Journal: Nature Communications

    Article Title: Engineering of high-precision base editors for site-specific single nucleotide replacement

    doi: 10.1038/s41467-018-08034-8

    Figure Lengend Snippet: Rigid linkers narrow the width of the editing window of BE3. a Protospacers and PAM (blue) sequences of the genomic loci tested, with the target Cs shown in red. Subscript numbers indicate the positions of the cytidines relative to the PAM. C-to-T editing at any of the indicated Cs inactivates the Can1 transporter and thus causes resistance to canavanine 17 . b Editing efficiency and specificity of the base editors tested as determined by canavanine selection. The x -axis represents the target Cs within the protospacers. The y -axis shows their C-to-T editing frequency (see Methods and Supplementary Figure 1 ). Sequences of canavanine-resistant mutants aligned with the corresponding reference sequences are shown in Supplementary Figure 3 . Values and error bars represent the mean and standard deviation of three independent biological replicates. Source data are provided as a Source Data file

    Article Snippet: The cells were incubated for 20 h prior to plating on YPAD rich or SC media plates without arginine but with 60 mg/mL l -canavanine (Sigma).

    Techniques: Selection, Standard Deviation

    Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.

    Journal: Emerging Microbes & Infections

    Article Title: Comparative genomic analysis of two emergent human adenovirus type 14 respiratory pathogen isolates in China reveals similar yet divergent genomes

    doi: 10.1038/emi.2017.78

    Figure Lengend Snippet: Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.

    Article Snippet: Genome type identification In silico restriction maps of available genome sequences from all HAdV-14 strains CHN/GZ01/2010, 303600, CHN/BJ430, CHN2012, and prototype de Wit were generated using the Vector NTI 10.3.0 software (Invitrogen Corp.; San Diego, CA, USA) as described in earlier studies., , These included profiles from restriction enzymes Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal l, Sma I, Xba I, and Xho I (TaKaRa Corp.; China), all of which were chosen in order to be consistent with the original nomenclature system devised by Li and colleagues.

    Techniques: In Silico, Generated, Plasmid Preparation, Software

    Gel retardation analysis to evaluate the inhibitory effects of rhamnulose-1-phophate (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR

    Journal: Journal of Bacteriology

    Article Title: Regulation of the rhaEWRBMA Operon Involved in l-Rhamnose Catabolism through Two Transcriptional Factors, RhaR and CcpA, in Bacillus subtilis

    doi: 10.1128/JB.00856-15

    Figure Lengend Snippet: Gel retardation analysis to evaluate the inhibitory effects of rhamnulose-1-phophate (A) and the phosphorylated rhamnulose produced by the His-RhaB-catalyzed reaction (B) on the DNA binding of RhaR. The 32 P-labeled P rhaEW probe (0.02 pmol) and the RhaR

    Article Snippet: By measuring the absorbance of the colored mixture at 545 nm, the amount of ketoses converted from rhamnose was quantified, based on the molar extinction coefficient at 545 nm (1,530 M−1 cm−1 ) determined by using l -rhamnulose (Sigma-Aldrich), which was dissolved at various concentrations with 50 mM Tris-Cl buffer (pH 8.0) plus 1 mM MnCl2 , and colored by the same treatment for spectrophotometric measurement.

    Techniques: Electrophoretic Mobility Shift Assay, Produced, Binding Assay, Labeling

    Treatment with L-Kynurenine fails to inhibit in vivo T CD8 responses to T Ag. WT mice were treated with L-Kyn (30 mg total as described in Materials and Methods) or PBS, and injected i.p. with C57SV cells. Nine days later, splenic T CD8 were examined for IFN-γ accumulation following restimulation with C57SV cells or synthetic peptides corresponding to T Ag epitopes. T Ag-specific T CD8 frequencies were determined after subtracting background and expressed as mean ± SEM of 6 L-Kyn-treated and 8 vehicle-treated mice (A). These values were used to calculate the absolute number of T Ag-specific T CD8 present within each spleen (B).

    Journal: PLoS ONE

    Article Title: Suppression of Immunodominant Antitumor and Antiviral CD8+ T Cell Responses by Indoleamine 2,3-Dioxygenase

    doi: 10.1371/journal.pone.0090439

    Figure Lengend Snippet: Treatment with L-Kynurenine fails to inhibit in vivo T CD8 responses to T Ag. WT mice were treated with L-Kyn (30 mg total as described in Materials and Methods) or PBS, and injected i.p. with C57SV cells. Nine days later, splenic T CD8 were examined for IFN-γ accumulation following restimulation with C57SV cells or synthetic peptides corresponding to T Ag epitopes. T Ag-specific T CD8 frequencies were determined after subtracting background and expressed as mean ± SEM of 6 L-Kyn-treated and 8 vehicle-treated mice (A). These values were used to calculate the absolute number of T Ag-specific T CD8 present within each spleen (B).

    Article Snippet: L-kynurenine (L-Kyn), also from Sigma, was dissolved at 10 mg/mL in PBS shortly before injection.

    Techniques: In Vivo, Mouse Assay, Injection