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mneongreen coding sequence  (Addgene inc)


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    Addgene inc mneongreen coding sequence
    BteA-induced cell death is characterized by fragmentation of the endoplasmatic reticulum and mitochondrial networks. ( A ) Time lapse analysis of morphological changes and plasma membrane permeabilization in Hela cells. Hela cells were infected with B. bronchiseptica wild-type ( Bb WT) and Bb Δ bteA mutant ( Bb Δ bteA ), expressing the fluorescent protein <t>mNeonGreen,</t> at MOI of 10:1 in the presence of propidium iodide (5 µg/mL). A sequence of time lapse images is shown. Bright field, gray; bacteria, cyan; propidium iodide, magenta. Scale bar, 20 µm. Data are representative of three independent experiments. ( B and C ) Comparison of B. bronchiseptica and B. pertussis cytotoxicity against HeLa cells. HeLa cells were infected with B. bronchiseptica ( B ) or B. pertussis ( C ) wild-type strains and their mutant derivatives at the indicated MOI. Plasma membrane permeabilization was determined using the fluorescent DNA binding dye CellTox Green. For B. bronchiseptica , infections were conducted both in the presence (gly+) and absence of glycine (5 mM). Asterisks indicate a statistically significant difference ( P < 0.05, unpaired two-tailed t -test) between Bb WT and Bb WT gly+ infection samples at 4 h. Data represent the mean ± SEM of a representative experiment out of 2 performed in technical triplicate. ( D ) Visualization of cellular structures. Hela cells were transfected to express fluorescent proteins tagged with localization signals for endoplasmatic reticulum (ER) and mitochondria (mito). One hour after infection with Bb WT and Bb Δ bteA mutant at MOI 50:1 , cells were fixed and analyzed by fluorescence imaging. ER, yellow; mitrochondria, magenta. Scale bar, 20 µm. The shown micrographs are representative of two independent experiments from which the organelle morphology was scored. Analysis was performed on at least 100 cells per experiment and condition and is plotted as morphology % ± SEM. Asterisks indicate statistically significant differences ( P < 0.05, unpaired two-tailed t -test) between the % of fragmented ER or mito compared with uninfected cells.
    Mneongreen Coding Sequence, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mneongreen+coding+sequence/4xmts-mNeonGreen+(Plasmid+%2398876)/pmc11633230-207-22-30
    Average 93 stars, based on 11 article reviews
    mneongreen coding sequence - by Bioz Stars, 2026-11
    93/100 stars

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    1) Product Images from "The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis"

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis

    Journal: mBio

    doi: 10.1128/mbio.01925-24

    BteA-induced cell death is characterized by fragmentation of the endoplasmatic reticulum and mitochondrial networks. ( A ) Time lapse analysis of morphological changes and plasma membrane permeabilization in Hela cells. Hela cells were infected with B. bronchiseptica wild-type ( Bb WT) and Bb Δ bteA mutant ( Bb Δ bteA ), expressing the fluorescent protein mNeonGreen, at MOI of 10:1 in the presence of propidium iodide (5 µg/mL). A sequence of time lapse images is shown. Bright field, gray; bacteria, cyan; propidium iodide, magenta. Scale bar, 20 µm. Data are representative of three independent experiments. ( B and C ) Comparison of B. bronchiseptica and B. pertussis cytotoxicity against HeLa cells. HeLa cells were infected with B. bronchiseptica ( B ) or B. pertussis ( C ) wild-type strains and their mutant derivatives at the indicated MOI. Plasma membrane permeabilization was determined using the fluorescent DNA binding dye CellTox Green. For B. bronchiseptica , infections were conducted both in the presence (gly+) and absence of glycine (5 mM). Asterisks indicate a statistically significant difference ( P < 0.05, unpaired two-tailed t -test) between Bb WT and Bb WT gly+ infection samples at 4 h. Data represent the mean ± SEM of a representative experiment out of 2 performed in technical triplicate. ( D ) Visualization of cellular structures. Hela cells were transfected to express fluorescent proteins tagged with localization signals for endoplasmatic reticulum (ER) and mitochondria (mito). One hour after infection with Bb WT and Bb Δ bteA mutant at MOI 50:1 , cells were fixed and analyzed by fluorescence imaging. ER, yellow; mitrochondria, magenta. Scale bar, 20 µm. The shown micrographs are representative of two independent experiments from which the organelle morphology was scored. Analysis was performed on at least 100 cells per experiment and condition and is plotted as morphology % ± SEM. Asterisks indicate statistically significant differences ( P < 0.05, unpaired two-tailed t -test) between the % of fragmented ER or mito compared with uninfected cells.
    Figure Legend Snippet: BteA-induced cell death is characterized by fragmentation of the endoplasmatic reticulum and mitochondrial networks. ( A ) Time lapse analysis of morphological changes and plasma membrane permeabilization in Hela cells. Hela cells were infected with B. bronchiseptica wild-type ( Bb WT) and Bb Δ bteA mutant ( Bb Δ bteA ), expressing the fluorescent protein mNeonGreen, at MOI of 10:1 in the presence of propidium iodide (5 µg/mL). A sequence of time lapse images is shown. Bright field, gray; bacteria, cyan; propidium iodide, magenta. Scale bar, 20 µm. Data are representative of three independent experiments. ( B and C ) Comparison of B. bronchiseptica and B. pertussis cytotoxicity against HeLa cells. HeLa cells were infected with B. bronchiseptica ( B ) or B. pertussis ( C ) wild-type strains and their mutant derivatives at the indicated MOI. Plasma membrane permeabilization was determined using the fluorescent DNA binding dye CellTox Green. For B. bronchiseptica , infections were conducted both in the presence (gly+) and absence of glycine (5 mM). Asterisks indicate a statistically significant difference ( P < 0.05, unpaired two-tailed t -test) between Bb WT and Bb WT gly+ infection samples at 4 h. Data represent the mean ± SEM of a representative experiment out of 2 performed in technical triplicate. ( D ) Visualization of cellular structures. Hela cells were transfected to express fluorescent proteins tagged with localization signals for endoplasmatic reticulum (ER) and mitochondria (mito). One hour after infection with Bb WT and Bb Δ bteA mutant at MOI 50:1 , cells were fixed and analyzed by fluorescence imaging. ER, yellow; mitrochondria, magenta. Scale bar, 20 µm. The shown micrographs are representative of two independent experiments from which the organelle morphology was scored. Analysis was performed on at least 100 cells per experiment and condition and is plotted as morphology % ± SEM. Asterisks indicate statistically significant differences ( P < 0.05, unpaired two-tailed t -test) between the % of fragmented ER or mito compared with uninfected cells.

    Techniques Used: Clinical Proteomics, Membrane, Infection, Mutagenesis, Expressing, Sequencing, Bacteria, Comparison, Binding Assay, CellTox Assay, Two Tailed Test, Transfection, Fluorescence, Imaging

    Related Articles

    Sequencing:

    Article Title: Pulses of RhoA signaling stimulate actin polymerization and flow in protrusions to drive collective cell migration.
    Article Snippet: .. For the cxcr4b:Myl12.1-mNeonGreen, themyl12.1 coding sequence was amplified from the targeting plasmid pUC19-homologyarm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm38 and the mNeonGreen coding sequence was amplified from mNeonGreenmTurquoise2 plasmid (Addgene #98886).83 The full name of this transgenic line is TgBAC(cxcr4b:Myl12.1-mNeonGreen)-p3 and the transgenesis marker is myl7:mScarlet. ..

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354–1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. All constructs were verified by DNA sequencing (Eurofins Genomics).

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354 – 1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Article Title: Pulses of RhoA Signaling Stimulate Actin Polymerization and Flow in Protrusions to Drive Collective Cell Migration
    Article Snippet: .. The myl12.1 coding sequence was amplified from the targeting plasmid pUC19-homology-arm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm ( ) and the mNeonGreen coding sequence was amplified from mNeonGreen-mTurquoise2 plasmid (Addgene #98886) ( ). ..

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System.
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent).

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis.
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354–1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Amplification:

    Article Title: Pulses of RhoA signaling stimulate actin polymerization and flow in protrusions to drive collective cell migration.
    Article Snippet: .. For the cxcr4b:Myl12.1-mNeonGreen, themyl12.1 coding sequence was amplified from the targeting plasmid pUC19-homologyarm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm38 and the mNeonGreen coding sequence was amplified from mNeonGreenmTurquoise2 plasmid (Addgene #98886).83 The full name of this transgenic line is TgBAC(cxcr4b:Myl12.1-mNeonGreen)-p3 and the transgenesis marker is myl7:mScarlet. ..

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354–1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. All constructs were verified by DNA sequencing (Eurofins Genomics).

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354 – 1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Article Title: Pulses of RhoA Signaling Stimulate Actin Polymerization and Flow in Protrusions to Drive Collective Cell Migration
    Article Snippet: .. The myl12.1 coding sequence was amplified from the targeting plasmid pUC19-homology-arm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm ( ) and the mNeonGreen coding sequence was amplified from mNeonGreen-mTurquoise2 plasmid (Addgene #98886) ( ). ..

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System.
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent).

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis.
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354–1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Plasmid Preparation:

    Article Title: Pulses of RhoA signaling stimulate actin polymerization and flow in protrusions to drive collective cell migration.
    Article Snippet: .. For the cxcr4b:Myl12.1-mNeonGreen, themyl12.1 coding sequence was amplified from the targeting plasmid pUC19-homologyarm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm38 and the mNeonGreen coding sequence was amplified from mNeonGreenmTurquoise2 plasmid (Addgene #98886).83 The full name of this transgenic line is TgBAC(cxcr4b:Myl12.1-mNeonGreen)-p3 and the transgenesis marker is myl7:mScarlet. ..

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis
    Article Snippet: .. The GroES promotor (391 nt, NC_002927.3, 1 041 354–1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA). ..

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. All constructs were verified by DNA sequencing (Eurofins Genomics).

    Article Title: Pulses of RhoA Signaling Stimulate Actin Polymerization and Flow in Protrusions to Drive Collective Cell Migration
    Article Snippet: .. The myl12.1 coding sequence was amplified from the targeting plasmid pUC19-homology-arm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm ( ) and the mNeonGreen coding sequence was amplified from mNeonGreen-mTurquoise2 plasmid (Addgene #98886) ( ). ..

    Transgenic Assay:

    Article Title: Pulses of RhoA signaling stimulate actin polymerization and flow in protrusions to drive collective cell migration.
    Article Snippet: .. For the cxcr4b:Myl12.1-mNeonGreen, themyl12.1 coding sequence was amplified from the targeting plasmid pUC19-homologyarm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm38 and the mNeonGreen coding sequence was amplified from mNeonGreenmTurquoise2 plasmid (Addgene #98886).83 The full name of this transgenic line is TgBAC(cxcr4b:Myl12.1-mNeonGreen)-p3 and the transgenesis marker is myl7:mScarlet. ..

    Marker:

    Article Title: Pulses of RhoA signaling stimulate actin polymerization and flow in protrusions to drive collective cell migration.
    Article Snippet: .. For the cxcr4b:Myl12.1-mNeonGreen, themyl12.1 coding sequence was amplified from the targeting plasmid pUC19-homologyarm-myl12.1-mScarlet-FRT-galK-FRT-homology-arm38 and the mNeonGreen coding sequence was amplified from mNeonGreenmTurquoise2 plasmid (Addgene #98886).83 The full name of this transgenic line is TgBAC(cxcr4b:Myl12.1-mNeonGreen)-p3 and the transgenesis marker is myl7:mScarlet. ..

    Synthesized:

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. All constructs were verified by DNA sequencing (Eurofins Genomics).

    Article Title: BopN is a Gatekeeper of the Bordetella Type III Secretion System.
    Article Snippet: PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent). .. The LgBit coding sequence was synthesized as GeneArt strings fragments (Invitrogen, Thermo Fisher Scientific), and mNeonGreen coding sequence was amplified from mNeonGreen-Rab7 vector (Addgene #129603). .. PCR amplifications were performed from chromosomal DNA of B. pertussis Tohama I or B. bronchiseptica RB50 using Herculase II Phusion DNA polymerase (Agilent).



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    BteA-induced cell death is characterized by fragmentation of the endoplasmatic reticulum and mitochondrial networks. ( A ) Time lapse analysis of morphological changes and plasma membrane permeabilization in Hela cells. Hela cells were infected with B. bronchiseptica wild-type ( Bb WT) and Bb Δ bteA mutant ( Bb Δ bteA ), expressing the fluorescent protein <t>mNeonGreen,</t> at MOI of 10:1 in the presence of propidium iodide (5 µg/mL). A sequence of time lapse images is shown. Bright field, gray; bacteria, cyan; propidium iodide, magenta. Scale bar, 20 µm. Data are representative of three independent experiments. ( B and C ) Comparison of B. bronchiseptica and B. pertussis cytotoxicity against HeLa cells. HeLa cells were infected with B. bronchiseptica ( B ) or B. pertussis ( C ) wild-type strains and their mutant derivatives at the indicated MOI. Plasma membrane permeabilization was determined using the fluorescent DNA binding dye CellTox Green. For B. bronchiseptica , infections were conducted both in the presence (gly+) and absence of glycine (5 mM). Asterisks indicate a statistically significant difference ( P < 0.05, unpaired two-tailed t -test) between Bb WT and Bb WT gly+ infection samples at 4 h. Data represent the mean ± SEM of a representative experiment out of 2 performed in technical triplicate. ( D ) Visualization of cellular structures. Hela cells were transfected to express fluorescent proteins tagged with localization signals for endoplasmatic reticulum (ER) and mitochondria (mito). One hour after infection with Bb WT and Bb Δ bteA mutant at MOI 50:1 , cells were fixed and analyzed by fluorescence imaging. ER, yellow; mitrochondria, magenta. Scale bar, 20 µm. The shown micrographs are representative of two independent experiments from which the organelle morphology was scored. Analysis was performed on at least 100 cells per experiment and condition and is plotted as morphology % ± SEM. Asterisks indicate statistically significant differences ( P < 0.05, unpaired two-tailed t -test) between the % of fragmented ER or mito compared with uninfected cells.
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    Image Search Results


    BteA-induced cell death is characterized by fragmentation of the endoplasmatic reticulum and mitochondrial networks. ( A ) Time lapse analysis of morphological changes and plasma membrane permeabilization in Hela cells. Hela cells were infected with B. bronchiseptica wild-type ( Bb WT) and Bb Δ bteA mutant ( Bb Δ bteA ), expressing the fluorescent protein mNeonGreen, at MOI of 10:1 in the presence of propidium iodide (5 µg/mL). A sequence of time lapse images is shown. Bright field, gray; bacteria, cyan; propidium iodide, magenta. Scale bar, 20 µm. Data are representative of three independent experiments. ( B and C ) Comparison of B. bronchiseptica and B. pertussis cytotoxicity against HeLa cells. HeLa cells were infected with B. bronchiseptica ( B ) or B. pertussis ( C ) wild-type strains and their mutant derivatives at the indicated MOI. Plasma membrane permeabilization was determined using the fluorescent DNA binding dye CellTox Green. For B. bronchiseptica , infections were conducted both in the presence (gly+) and absence of glycine (5 mM). Asterisks indicate a statistically significant difference ( P < 0.05, unpaired two-tailed t -test) between Bb WT and Bb WT gly+ infection samples at 4 h. Data represent the mean ± SEM of a representative experiment out of 2 performed in technical triplicate. ( D ) Visualization of cellular structures. Hela cells were transfected to express fluorescent proteins tagged with localization signals for endoplasmatic reticulum (ER) and mitochondria (mito). One hour after infection with Bb WT and Bb Δ bteA mutant at MOI 50:1 , cells were fixed and analyzed by fluorescence imaging. ER, yellow; mitrochondria, magenta. Scale bar, 20 µm. The shown micrographs are representative of two independent experiments from which the organelle morphology was scored. Analysis was performed on at least 100 cells per experiment and condition and is plotted as morphology % ± SEM. Asterisks indicate statistically significant differences ( P < 0.05, unpaired two-tailed t -test) between the % of fragmented ER or mito compared with uninfected cells.

    Journal: mBio

    Article Title: The Bordetella effector protein BteA induces host cell death by disruption of calcium homeostasis

    doi: 10.1128/mbio.01925-24

    Figure Lengend Snippet: BteA-induced cell death is characterized by fragmentation of the endoplasmatic reticulum and mitochondrial networks. ( A ) Time lapse analysis of morphological changes and plasma membrane permeabilization in Hela cells. Hela cells were infected with B. bronchiseptica wild-type ( Bb WT) and Bb Δ bteA mutant ( Bb Δ bteA ), expressing the fluorescent protein mNeonGreen, at MOI of 10:1 in the presence of propidium iodide (5 µg/mL). A sequence of time lapse images is shown. Bright field, gray; bacteria, cyan; propidium iodide, magenta. Scale bar, 20 µm. Data are representative of three independent experiments. ( B and C ) Comparison of B. bronchiseptica and B. pertussis cytotoxicity against HeLa cells. HeLa cells were infected with B. bronchiseptica ( B ) or B. pertussis ( C ) wild-type strains and their mutant derivatives at the indicated MOI. Plasma membrane permeabilization was determined using the fluorescent DNA binding dye CellTox Green. For B. bronchiseptica , infections were conducted both in the presence (gly+) and absence of glycine (5 mM). Asterisks indicate a statistically significant difference ( P < 0.05, unpaired two-tailed t -test) between Bb WT and Bb WT gly+ infection samples at 4 h. Data represent the mean ± SEM of a representative experiment out of 2 performed in technical triplicate. ( D ) Visualization of cellular structures. Hela cells were transfected to express fluorescent proteins tagged with localization signals for endoplasmatic reticulum (ER) and mitochondria (mito). One hour after infection with Bb WT and Bb Δ bteA mutant at MOI 50:1 , cells were fixed and analyzed by fluorescence imaging. ER, yellow; mitrochondria, magenta. Scale bar, 20 µm. The shown micrographs are representative of two independent experiments from which the organelle morphology was scored. Analysis was performed on at least 100 cells per experiment and condition and is plotted as morphology % ± SEM. Asterisks indicate statistically significant differences ( P < 0.05, unpaired two-tailed t -test) between the % of fragmented ER or mito compared with uninfected cells.

    Article Snippet: The GroES promotor (391 nt, NC_002927.3, 1 041 354–1 041 744) was amplified from chromosomal DNA of B. bronchiseptica RB50, whereas the mNeonGreen coding sequence was amplified from 4xmts-mNeonGreen vector (Addgene item # 98876) using Herculase II Phusion DNA polymerase (Agilent, USA).

    Techniques: Clinical Proteomics, Membrane, Infection, Mutagenesis, Expressing, Sequencing, Bacteria, Comparison, Binding Assay, CellTox Assay, Two Tailed Test, Transfection, Fluorescence, Imaging