e coli dna polymerase Thermo Fisher Search Results


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  • 95
    New England Biolabs polymerase
    Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 833 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher dna polymerase i e coli
    Nicking enzyme-mediated labeling of open chromatin. a Nicking of crosslinked chromatin using varying amounts of Nt.CviPII. A 1% agarose gel showing differential nicking of HCT116 genomic <t>DNA</t> based on amount of nicking enzyme (10 U, 5 U, 2.5 U, 1 U, 0.3 U, 0 U). M is a DNA molecular weight ladder. b Open chromatin labeling in fixed HeLa cells using dNTPs supplemented with TexasRed-dATP. Top panel : labeling reaction performed in the presence of Nt.CviPII and DNA polymerase I. Middle panel : labeling reaction performed in the presence of DNA <t>polymerase</t> I only. Bottom panel : labeling reaction performed in the absence of Nt.CviPII and DNA polymerase I. TexasRed-dATP was included in all reactions. DNA staining was performed using DAPI ( blue ) and TexasRed stain ( red ) represents labeled OCSs. Magenta stain ( Merge ) represents the colocalization. c Labeling efficiency of OCSs in all three assayed conditions. The y-axis represents the ratio of the intensity of the red pixels to the intensity of the blue pixels ( OCS labeling efficiency ). d Dot blot showing labeling of open chromatin by Nt.CviPII nicking enzyme in both native and formaldehyde-fixed HCT116 cells. The level of labeling was revealed using HRP-conjugated goat anti-biotin antibody
    Dna Polymerase I E Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Thermo Fisher escherichia coli dna polymerase
    Nicking enzyme-mediated labeling of open chromatin. a Nicking of crosslinked chromatin using varying amounts of Nt.CviPII. A 1% agarose gel showing differential nicking of HCT116 genomic <t>DNA</t> based on amount of nicking enzyme (10 U, 5 U, 2.5 U, 1 U, 0.3 U, 0 U). M is a DNA molecular weight ladder. b Open chromatin labeling in fixed HeLa cells using dNTPs supplemented with TexasRed-dATP. Top panel : labeling reaction performed in the presence of Nt.CviPII and DNA polymerase I. Middle panel : labeling reaction performed in the presence of DNA <t>polymerase</t> I only. Bottom panel : labeling reaction performed in the absence of Nt.CviPII and DNA polymerase I. TexasRed-dATP was included in all reactions. DNA staining was performed using DAPI ( blue ) and TexasRed stain ( red ) represents labeled OCSs. Magenta stain ( Merge ) represents the colocalization. c Labeling efficiency of OCSs in all three assayed conditions. The y-axis represents the ratio of the intensity of the red pixels to the intensity of the blue pixels ( OCS labeling efficiency ). d Dot blot showing labeling of open chromatin by Nt.CviPII nicking enzyme in both native and formaldehyde-fixed HCT116 cells. The level of labeling was revealed using HRP-conjugated goat anti-biotin antibody
    Escherichia Coli Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher e coli dna
    Promotion of plasma cell differentiation and Ig production in lupus B cells by <t>ALD-DNA.</t> Splenic B cells from 16- to 20-week-old MRL/lpr mice and 12-week-old MRL +/+ mice were cultured in media alone, or in the presence of ALD-DNA (50 µg/ml), LPS (100 ng/ml), or both for different periods as indicated below. ( A ) After 72 h stimulation, viable cells were analyzed for B220 and CD138 surface expression by flow cytometry. Representative dot plots showed the frequencies of plasmablasts (B220 + CD138 hi , lower panels, MRL/lpr, n = 6) or plasma cells (B220 − CD138 + , upper panels, MRL +/+ , n = 6) generated after each treatment. ( B ) Expression of Blimp-1 and XBP1 mRNA from lupus B cells cultured for 96 h was examined by real-time PCR. Results are expressed as the fold induction of gene transcription relative to their untreated controls (means ±SEM, n = 6). ( C ) Ig production in cell culture supernatants harvested on day 6 was determined by ELISA. Bars represent the means ±SEM of three independent experiments (n = 6). * P
    E Coli Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 164 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher escherichia coli dh10b
    Promotion of plasma cell differentiation and Ig production in lupus B cells by <t>ALD-DNA.</t> Splenic B cells from 16- to 20-week-old MRL/lpr mice and 12-week-old MRL +/+ mice were cultured in media alone, or in the presence of ALD-DNA (50 µg/ml), LPS (100 ng/ml), or both for different periods as indicated below. ( A ) After 72 h stimulation, viable cells were analyzed for B220 and CD138 surface expression by flow cytometry. Representative dot plots showed the frequencies of plasmablasts (B220 + CD138 hi , lower panels, MRL/lpr, n = 6) or plasma cells (B220 − CD138 + , upper panels, MRL +/+ , n = 6) generated after each treatment. ( B ) Expression of Blimp-1 and XBP1 mRNA from lupus B cells cultured for 96 h was examined by real-time PCR. Results are expressed as the fold induction of gene transcription relative to their untreated controls (means ±SEM, n = 6). ( C ) Ig production in cell culture supernatants harvested on day 6 was determined by ELISA. Bars represent the means ±SEM of three independent experiments (n = 6). * P
    Escherichia Coli Dh10b, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 576 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher top10 escherichia coli
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Top10 Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 350 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher top10 competent escherichia coli
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Top10 Competent Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher e coli dh5α
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    E Coli Dh5α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 2953 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher escherichia coli
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 5728 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher e coli total rna
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    E Coli Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher competent escherichia coli cells
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Competent Escherichia Coli Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher escherichia coli strain top10
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Escherichia Coli Strain Top10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 518 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher escherichia coli poly
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Escherichia Coli Poly, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher pfx50 dna polymerase
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Pfx50 Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 320 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher elongase dna polymerase
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Elongase Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher accuprime pfx dna polymerase
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Accuprime Pfx Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2231 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher escherichia coli dh5a competent cells
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Escherichia Coli Dh5a Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher e coli top10 cells
    Competition assays and G. mellonella killing models in HLCRMs. a Fitness of full-length mcr-1 , calalytically inactivated mcr-1 (E246A), and mcr-1 soluble domain. Fitness of blaTEM-1b as a negative control. b Relative fitness of wild-type mcr-1 -positive strains and their derivatives. c Relative fitness of mcr-1/ pBAD-positive E.coli <t>TOP10</t> strains and their derivatives. In all fitness figures, error bars represent the SD ( n = 6). The differences in fitness were tested using non-parametric Mann–Whitney test, * indicates 0.01
    E Coli Top10 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1873 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Nicking enzyme-mediated labeling of open chromatin. a Nicking of crosslinked chromatin using varying amounts of Nt.CviPII. A 1% agarose gel showing differential nicking of HCT116 genomic DNA based on amount of nicking enzyme (10 U, 5 U, 2.5 U, 1 U, 0.3 U, 0 U). M is a DNA molecular weight ladder. b Open chromatin labeling in fixed HeLa cells using dNTPs supplemented with TexasRed-dATP. Top panel : labeling reaction performed in the presence of Nt.CviPII and DNA polymerase I. Middle panel : labeling reaction performed in the presence of DNA polymerase I only. Bottom panel : labeling reaction performed in the absence of Nt.CviPII and DNA polymerase I. TexasRed-dATP was included in all reactions. DNA staining was performed using DAPI ( blue ) and TexasRed stain ( red ) represents labeled OCSs. Magenta stain ( Merge ) represents the colocalization. c Labeling efficiency of OCSs in all three assayed conditions. The y-axis represents the ratio of the intensity of the red pixels to the intensity of the blue pixels ( OCS labeling efficiency ). d Dot blot showing labeling of open chromatin by Nt.CviPII nicking enzyme in both native and formaldehyde-fixed HCT116 cells. The level of labeling was revealed using HRP-conjugated goat anti-biotin antibody

    Journal: Genome Biology

    Article Title: NicE-seq: high resolution open chromatin profiling

    doi: 10.1186/s13059-017-1247-6

    Figure Lengend Snippet: Nicking enzyme-mediated labeling of open chromatin. a Nicking of crosslinked chromatin using varying amounts of Nt.CviPII. A 1% agarose gel showing differential nicking of HCT116 genomic DNA based on amount of nicking enzyme (10 U, 5 U, 2.5 U, 1 U, 0.3 U, 0 U). M is a DNA molecular weight ladder. b Open chromatin labeling in fixed HeLa cells using dNTPs supplemented with TexasRed-dATP. Top panel : labeling reaction performed in the presence of Nt.CviPII and DNA polymerase I. Middle panel : labeling reaction performed in the presence of DNA polymerase I only. Bottom panel : labeling reaction performed in the absence of Nt.CviPII and DNA polymerase I. TexasRed-dATP was included in all reactions. DNA staining was performed using DAPI ( blue ) and TexasRed stain ( red ) represents labeled OCSs. Magenta stain ( Merge ) represents the colocalization. c Labeling efficiency of OCSs in all three assayed conditions. The y-axis represents the ratio of the intensity of the red pixels to the intensity of the blue pixels ( OCS labeling efficiency ). d Dot blot showing labeling of open chromatin by Nt.CviPII nicking enzyme in both native and formaldehyde-fixed HCT116 cells. The level of labeling was revealed using HRP-conjugated goat anti-biotin antibody

    Article Snippet: Open chromatin DNA was labeled with biotin by incubating the nuclei in the presence of 2.5 U of Nt.CviPII (NEB R0626S), 10 U of DNA polymerase I (M0209S) and 30 μM of each dNTP including 6 μM of biotin-14-dATP (Invitrogen, 19524-016) and 6 μM of biotin-16-dCTP (ChemCyte, CC-6003-1) in 200 μL of 1× NEB buffer 2.

    Techniques: Labeling, Agarose Gel Electrophoresis, Molecular Weight, Staining, Dot Blot

    Promotion of plasma cell differentiation and Ig production in lupus B cells by ALD-DNA. Splenic B cells from 16- to 20-week-old MRL/lpr mice and 12-week-old MRL +/+ mice were cultured in media alone, or in the presence of ALD-DNA (50 µg/ml), LPS (100 ng/ml), or both for different periods as indicated below. ( A ) After 72 h stimulation, viable cells were analyzed for B220 and CD138 surface expression by flow cytometry. Representative dot plots showed the frequencies of plasmablasts (B220 + CD138 hi , lower panels, MRL/lpr, n = 6) or plasma cells (B220 − CD138 + , upper panels, MRL +/+ , n = 6) generated after each treatment. ( B ) Expression of Blimp-1 and XBP1 mRNA from lupus B cells cultured for 96 h was examined by real-time PCR. Results are expressed as the fold induction of gene transcription relative to their untreated controls (means ±SEM, n = 6). ( C ) Ig production in cell culture supernatants harvested on day 6 was determined by ELISA. Bars represent the means ±SEM of three independent experiments (n = 6). * P

    Journal: PLoS ONE

    Article Title: Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production

    doi: 10.1371/journal.pone.0109095

    Figure Lengend Snippet: Promotion of plasma cell differentiation and Ig production in lupus B cells by ALD-DNA. Splenic B cells from 16- to 20-week-old MRL/lpr mice and 12-week-old MRL +/+ mice were cultured in media alone, or in the presence of ALD-DNA (50 µg/ml), LPS (100 ng/ml), or both for different periods as indicated below. ( A ) After 72 h stimulation, viable cells were analyzed for B220 and CD138 surface expression by flow cytometry. Representative dot plots showed the frequencies of plasmablasts (B220 + CD138 hi , lower panels, MRL/lpr, n = 6) or plasma cells (B220 − CD138 + , upper panels, MRL +/+ , n = 6) generated after each treatment. ( B ) Expression of Blimp-1 and XBP1 mRNA from lupus B cells cultured for 96 h was examined by real-time PCR. Results are expressed as the fold induction of gene transcription relative to their untreated controls (means ±SEM, n = 6). ( C ) Ig production in cell culture supernatants harvested on day 6 was determined by ELISA. Bars represent the means ±SEM of three independent experiments (n = 6). * P

    Article Snippet: CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences).

    Techniques: Cell Differentiation, Mouse Assay, Cell Culture, Expressing, Flow Cytometry, Cytometry, Generated, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Up-regulation of CD86 and MHC class II expression in naïve B cells by ALD-DNA stimulation. ( A ) Naïve B cells were incubated with ALD-DNA (50 µg/ml) in the presence or absence of LPS ranging from 0.01 µg/mL to 10 µg/ml for 72 h. The numbers of CD86-expressing B cells after each treatment were determined by flow cytometry. ( B ) Naive B cells were stimulated with ALD-DNA or E. coli single-stranded (ss) DNA (both at 50 µg/ml) for 72 h. CD86 and MHC class II expression was determined by performing flow cytometry. Histogram plots show the mean fluorescence intensity (MFI) of these surface molecules. The data are representative of three independent experiments.

    Journal: PLoS ONE

    Article Title: Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production

    doi: 10.1371/journal.pone.0109095

    Figure Lengend Snippet: Up-regulation of CD86 and MHC class II expression in naïve B cells by ALD-DNA stimulation. ( A ) Naïve B cells were incubated with ALD-DNA (50 µg/ml) in the presence or absence of LPS ranging from 0.01 µg/mL to 10 µg/ml for 72 h. The numbers of CD86-expressing B cells after each treatment were determined by flow cytometry. ( B ) Naive B cells were stimulated with ALD-DNA or E. coli single-stranded (ss) DNA (both at 50 µg/ml) for 72 h. CD86 and MHC class II expression was determined by performing flow cytometry. Histogram plots show the mean fluorescence intensity (MFI) of these surface molecules. The data are representative of three independent experiments.

    Article Snippet: CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences).

    Techniques: Expressing, Incubation, Flow Cytometry, Cytometry, Fluorescence

    Enhancement of LPS-stimulated B cell survival and division by ALD-DNA. ( A ) CFSE-labeled B cells were stimulated with ALD-DNA (50 µg/ml) in the presence or absence of LPS (100 ng/ml) for 72 h. Division progression of B cells was determined by dilution of CFSE. Representative histogram plots show the percentage of proliferating (CFSE-low) B cells after each treatment (mean values ±SEM, n = 6). Shaded areas show the cell division peaks predicted by the ModFit software. ( B ) Flow cytometry dot plots showing the gate used to select living cells and eliminate cell debris, and showing the viability of B cells (mean values ±SEM, n = 11) under different culture conditions as described above for 72 h. ( C ) Nude mice were administered one intravenous injection of indicated stimuli at day 1 and an additional intraperitoneal injection of BrdU 24 h before sacrifice. After 72 h, spleens were collected, and B220 + B cells with incorporated BrdU combined with total DNA content (with 7-AAD) were analyzed by FACS. The region gates applied to the 7-AAD versus BrdU dot plot indicate the cell subsets that were resided in the S phase of the cell cycle. Results represent 6 mice/experimental group from two separate experiments.

    Journal: PLoS ONE

    Article Title: Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production

    doi: 10.1371/journal.pone.0109095

    Figure Lengend Snippet: Enhancement of LPS-stimulated B cell survival and division by ALD-DNA. ( A ) CFSE-labeled B cells were stimulated with ALD-DNA (50 µg/ml) in the presence or absence of LPS (100 ng/ml) for 72 h. Division progression of B cells was determined by dilution of CFSE. Representative histogram plots show the percentage of proliferating (CFSE-low) B cells after each treatment (mean values ±SEM, n = 6). Shaded areas show the cell division peaks predicted by the ModFit software. ( B ) Flow cytometry dot plots showing the gate used to select living cells and eliminate cell debris, and showing the viability of B cells (mean values ±SEM, n = 11) under different culture conditions as described above for 72 h. ( C ) Nude mice were administered one intravenous injection of indicated stimuli at day 1 and an additional intraperitoneal injection of BrdU 24 h before sacrifice. After 72 h, spleens were collected, and B220 + B cells with incorporated BrdU combined with total DNA content (with 7-AAD) were analyzed by FACS. The region gates applied to the 7-AAD versus BrdU dot plot indicate the cell subsets that were resided in the S phase of the cell cycle. Results represent 6 mice/experimental group from two separate experiments.

    Article Snippet: CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences).

    Techniques: Labeling, Software, Flow Cytometry, Cytometry, Mouse Assay, Injection, FACS

    Enhancement of CD40-stimulated naïve B cell proliferation by ALD-DNA. ( A ) Naïve B cells were pre-incubated for 20 min with polymyxin B (PMB; 20 µg/ml) before the addition of ALD-DNA (50 µg/ml), LPS (100 ng/ml), anti-CD40 (1 µg/ml), LPS (100 ng/ml) plus ALD-DNA (50 µg/ml), or anti-CD40 (1 µg/ml) plus ALD-DNA (50 µg/ml). After 72 h stimulation, cell proliferation was detected by performing BrdU incorporation assay using ELISA. Data are presented as the means ±SEM of three independent experiments. ( B ) CFSE-labeled naïve B cells were cultured in media containing ALD-DNA, anti-CD40, or both (at the concentrations as described above) for 72 h. Cell division was monitored by measuring the dilution of CFSE. Shaded areas show the cell division peaks predicted by the ModFit software. Results from one representative experiment of three are shown. ** P

    Journal: PLoS ONE

    Article Title: Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production

    doi: 10.1371/journal.pone.0109095

    Figure Lengend Snippet: Enhancement of CD40-stimulated naïve B cell proliferation by ALD-DNA. ( A ) Naïve B cells were pre-incubated for 20 min with polymyxin B (PMB; 20 µg/ml) before the addition of ALD-DNA (50 µg/ml), LPS (100 ng/ml), anti-CD40 (1 µg/ml), LPS (100 ng/ml) plus ALD-DNA (50 µg/ml), or anti-CD40 (1 µg/ml) plus ALD-DNA (50 µg/ml). After 72 h stimulation, cell proliferation was detected by performing BrdU incorporation assay using ELISA. Data are presented as the means ±SEM of three independent experiments. ( B ) CFSE-labeled naïve B cells were cultured in media containing ALD-DNA, anti-CD40, or both (at the concentrations as described above) for 72 h. Cell division was monitored by measuring the dilution of CFSE. Shaded areas show the cell division peaks predicted by the ModFit software. Results from one representative experiment of three are shown. ** P

    Article Snippet: CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences).

    Techniques: Incubation, BrdU Incorporation Assay, Enzyme-linked Immunosorbent Assay, Labeling, Cell Culture, Software

    Synergistic enhancement of LPS-driven IgG production in naive B cells by ALD-DNA stimulation. Naive B cells were cultured for 6 days in media containing ALD-DNA (50 µg/ml) with or without LPS (100 ng/ml). Cell culture supernatants were collected, and the levels of IgM and IgG were measured by ELISA. Bars represent the means ±SEM of three independent experiments (n = 5). ND (not detected). ** P

    Journal: PLoS ONE

    Article Title: Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production

    doi: 10.1371/journal.pone.0109095

    Figure Lengend Snippet: Synergistic enhancement of LPS-driven IgG production in naive B cells by ALD-DNA stimulation. Naive B cells were cultured for 6 days in media containing ALD-DNA (50 µg/ml) with or without LPS (100 ng/ml). Cell culture supernatants were collected, and the levels of IgM and IgG were measured by ELISA. Bars represent the means ±SEM of three independent experiments (n = 5). ND (not detected). ** P

    Article Snippet: CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences).

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

    Effect of ALD-DNA on CD138, Blimp-1, XBP1, and IL-6 expression in naïve B cells. Naïve B cells were cultured in media containing ALD-DNA (50 µg/ml), LPS (100 ng/ml), or both for different periods as described below. ( A ) After 72 h stimulation, Cells were analyzed for B220 and CD138 surface expression by flow cytometry. The number indicates the percentage of B220 − CD138 + cells in the gate. Similar results were obtained in three independent experiments. ( B ) Expression of Blimp-1 and XBP1 mRNA from B cells cultured for 96 hours was examined by real-time PCR. Fold induction relative to their untreated controls is shown (means ±SEM, n = 5). ( C ) Expression of IL-6 mRNA in B cells after 6 h stimulation was examined by real-time PCR. Fold induction relative to their untreated controls is shown (means ±SEM, n = 5); The amounts of IL-6 protein in the culture supernatants of B cells after 72 h stimulation were measured by ELISA (n = 6, a line linked two dots represents a pair observation). Up-regulation of IL-6 protein was undetectable in all samples after ALD-DNA treatment. * P

    Journal: PLoS ONE

    Article Title: Self DNA from Lymphocytes That Have Undergone Activation-Induced Cell Death Enhances Murine B Cell Proliferation and Antibody Production

    doi: 10.1371/journal.pone.0109095

    Figure Lengend Snippet: Effect of ALD-DNA on CD138, Blimp-1, XBP1, and IL-6 expression in naïve B cells. Naïve B cells were cultured in media containing ALD-DNA (50 µg/ml), LPS (100 ng/ml), or both for different periods as described below. ( A ) After 72 h stimulation, Cells were analyzed for B220 and CD138 surface expression by flow cytometry. The number indicates the percentage of B220 − CD138 + cells in the gate. Similar results were obtained in three independent experiments. ( B ) Expression of Blimp-1 and XBP1 mRNA from B cells cultured for 96 hours was examined by real-time PCR. Fold induction relative to their untreated controls is shown (means ±SEM, n = 5). ( C ) Expression of IL-6 mRNA in B cells after 6 h stimulation was examined by real-time PCR. Fold induction relative to their untreated controls is shown (means ±SEM, n = 5); The amounts of IL-6 protein in the culture supernatants of B cells after 72 h stimulation were measured by ELISA (n = 6, a line linked two dots represents a pair observation). Up-regulation of IL-6 protein was undetectable in all samples after ALD-DNA treatment. * P

    Article Snippet: CFSE-labeled B cells stimulated with ALD-DNA, E. coli DNA, LPS, anti-CD40, or different combinations of these stimuli for 3 days were stained with B220-APC (RA3-6B2, eBioscience) or CD138-APC (281-2, BD Biosciences).

    Techniques: Expressing, Cell Culture, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Journal: Clinical and Experimental Immunology

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer

    doi: 10.1111/cei.13183

    Figure Lengend Snippet: DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Article Snippet: Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol.

    Techniques: DNA Methylation Assay, Sequencing, Cell Culture, Recombinant, In Vitro, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Expressing, Clone Assay, Plasmid Preparation, Transformation Assay

    Competition assays and G. mellonella killing models in HLCRMs. a Fitness of full-length mcr-1 , calalytically inactivated mcr-1 (E246A), and mcr-1 soluble domain. Fitness of blaTEM-1b as a negative control. b Relative fitness of wild-type mcr-1 -positive strains and their derivatives. c Relative fitness of mcr-1/ pBAD-positive E.coli TOP10 strains and their derivatives. In all fitness figures, error bars represent the SD ( n = 6). The differences in fitness were tested using non-parametric Mann–Whitney test, * indicates 0.01

    Journal: Nature Communications

    Article Title: Balancing mcr-1 expression and bacterial survival is a delicate equilibrium between essential cellular defence mechanisms

    doi: 10.1038/s41467-017-02149-0

    Figure Lengend Snippet: Competition assays and G. mellonella killing models in HLCRMs. a Fitness of full-length mcr-1 , calalytically inactivated mcr-1 (E246A), and mcr-1 soluble domain. Fitness of blaTEM-1b as a negative control. b Relative fitness of wild-type mcr-1 -positive strains and their derivatives. c Relative fitness of mcr-1/ pBAD-positive E.coli TOP10 strains and their derivatives. In all fitness figures, error bars represent the SD ( n = 6). The differences in fitness were tested using non-parametric Mann–Whitney test, * indicates 0.01

    Article Snippet: All resultant plasmids were transformed into E. coli TOP10 cell (Invitrogen, UK), purified and its integrity confirmed by PCR, double restriction digestion and DNA sequencing.

    Techniques: Negative Control, MANN-WHITNEY

    TEM micrographs of untreated and treated E.coli . In a and b , TEM micrographs of untreated control cells ( E. coli TOP10 with pBAD minus mcr-1 , and E. coli TOP10 ( mcr-1 /pBAD) without l -arabinose induction, respectively); both cells are intact with a well-defined inner and outer membrane, and showed a highly homogeneous electron density in cytoplasm region ( d and e ). c TEM micrographs of mcr-1 overproducing cells; the damaging outer membrane and some completely lysed cells were observed ( f )

    Journal: Nature Communications

    Article Title: Balancing mcr-1 expression and bacterial survival is a delicate equilibrium between essential cellular defence mechanisms

    doi: 10.1038/s41467-017-02149-0

    Figure Lengend Snippet: TEM micrographs of untreated and treated E.coli . In a and b , TEM micrographs of untreated control cells ( E. coli TOP10 with pBAD minus mcr-1 , and E. coli TOP10 ( mcr-1 /pBAD) without l -arabinose induction, respectively); both cells are intact with a well-defined inner and outer membrane, and showed a highly homogeneous electron density in cytoplasm region ( d and e ). c TEM micrographs of mcr-1 overproducing cells; the damaging outer membrane and some completely lysed cells were observed ( f )

    Article Snippet: All resultant plasmids were transformed into E. coli TOP10 cell (Invitrogen, UK), purified and its integrity confirmed by PCR, double restriction digestion and DNA sequencing.

    Techniques: Transmission Electron Microscopy