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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
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    Images

    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

    other:

    Article Title: FidlTrack: high-fidelity structure-aware single particle tracking resolves intracellular molecular motion in organelles sensing APP processing
    Article Snippet: 250 , CMV_4xmts_mNeonGreen , mNeonGreen Mito , Mitochondria matrix targeted mNeonGreen , Fig. , Addgene #98876.

    In Vitro:

    Article Title: Microtubules coordinate mitochondria transport with myofibril morphogenesis during muscle development.
    Article Snippet: .. In vitro myofibers were transfected 3 days after plating using Lipofectamine 2000 (Thermo Fisher Scientific; 11668-019) following the manufacturer’s instructions: 1 μg of DNA per ibidi 35-mm dish of mMito-dendra2 construct (AddGene; Plasmid #55796) was used to label photoconvertible mitochondria, while 4xmts-mNeonGreen(AddGene; Plasmid #98876) was employed to label mitochondria for immunostaining; for kinesin-1 (KIF5B) depletion, cells were co-transfected with 0.5 μg of 4xmts-mNeonGreen (AddGene; Plasmid #98876) and 10 nM of KIF5B siRNA#1 (ID s68782) or siRNA#2 (ID s68781) (Thermo Fisher Scientific). .. Silencer Select Negative Control (Thermo Fisher Scientific; #4390843) was used as the control. e3 Developmental Cell 60, 2962–2975.e1–e5, November 3, 2025

    Transfection:

    Article Title: Microtubules coordinate mitochondria transport with myofibril morphogenesis during muscle development.
    Article Snippet: .. In vitro myofibers were transfected 3 days after plating using Lipofectamine 2000 (Thermo Fisher Scientific; 11668-019) following the manufacturer’s instructions: 1 μg of DNA per ibidi 35-mm dish of mMito-dendra2 construct (AddGene; Plasmid #55796) was used to label photoconvertible mitochondria, while 4xmts-mNeonGreen(AddGene; Plasmid #98876) was employed to label mitochondria for immunostaining; for kinesin-1 (KIF5B) depletion, cells were co-transfected with 0.5 μg of 4xmts-mNeonGreen (AddGene; Plasmid #98876) and 10 nM of KIF5B siRNA#1 (ID s68782) or siRNA#2 (ID s68781) (Thermo Fisher Scientific). .. Silencer Select Negative Control (Thermo Fisher Scientific; #4390843) was used as the control. e3 Developmental Cell 60, 2962–2975.e1–e5, November 3, 2025

    Construct:

    Article Title: Microtubules coordinate mitochondria transport with myofibril morphogenesis during muscle development.
    Article Snippet: .. In vitro myofibers were transfected 3 days after plating using Lipofectamine 2000 (Thermo Fisher Scientific; 11668-019) following the manufacturer’s instructions: 1 μg of DNA per ibidi 35-mm dish of mMito-dendra2 construct (AddGene; Plasmid #55796) was used to label photoconvertible mitochondria, while 4xmts-mNeonGreen(AddGene; Plasmid #98876) was employed to label mitochondria for immunostaining; for kinesin-1 (KIF5B) depletion, cells were co-transfected with 0.5 μg of 4xmts-mNeonGreen (AddGene; Plasmid #98876) and 10 nM of KIF5B siRNA#1 (ID s68782) or siRNA#2 (ID s68781) (Thermo Fisher Scientific). .. Silencer Select Negative Control (Thermo Fisher Scientific; #4390843) was used as the control. e3 Developmental Cell 60, 2962–2975.e1–e5, November 3, 2025

    Plasmid Preparation:

    Article Title: Microtubules coordinate mitochondria transport with myofibril morphogenesis during muscle development.
    Article Snippet: .. In vitro myofibers were transfected 3 days after plating using Lipofectamine 2000 (Thermo Fisher Scientific; 11668-019) following the manufacturer’s instructions: 1 μg of DNA per ibidi 35-mm dish of mMito-dendra2 construct (AddGene; Plasmid #55796) was used to label photoconvertible mitochondria, while 4xmts-mNeonGreen(AddGene; Plasmid #98876) was employed to label mitochondria for immunostaining; for kinesin-1 (KIF5B) depletion, cells were co-transfected with 0.5 μg of 4xmts-mNeonGreen (AddGene; Plasmid #98876) and 10 nM of KIF5B siRNA#1 (ID s68782) or siRNA#2 (ID s68781) (Thermo Fisher Scientific). .. Silencer Select Negative Control (Thermo Fisher Scientific; #4390843) was used as the control. e3 Developmental Cell 60, 2962–2975.e1–e5, November 3, 2025

    Article Title: The Role of the Tyrosine-Based Sorting Signals of the ORF3a Protein of SARS-CoV-2 in Intracellular Trafficking and Pathogenesis
    Article Snippet: .. To examine the co-localization of the ORF3a proteins with mitochondria, COS-7 cells were co-transfected with vectors expressing the ORF3a proteins and a vector expressing 4xmts-NeonGreen (mitochondria; Addgene, #98876). ..

    Article Title: The Role of the Tyrosine-Based Sorting Signals of the ORF3a Protein of SARS-CoV-2 in Intracellular Trafficking and Pathogenesis
    Article Snippet: .. To examine the co-localization of the ORF3a proteins with mitochondria, COS-7 cells were co-transfected with vectors expressing the ORF3a proteins and a vector expressing 4xmts-NeonGreen (mitochondria; Addgene, #98876). ..

    Immunostaining:

    Article Title: Microtubules coordinate mitochondria transport with myofibril morphogenesis during muscle development.
    Article Snippet: .. In vitro myofibers were transfected 3 days after plating using Lipofectamine 2000 (Thermo Fisher Scientific; 11668-019) following the manufacturer’s instructions: 1 μg of DNA per ibidi 35-mm dish of mMito-dendra2 construct (AddGene; Plasmid #55796) was used to label photoconvertible mitochondria, while 4xmts-mNeonGreen(AddGene; Plasmid #98876) was employed to label mitochondria for immunostaining; for kinesin-1 (KIF5B) depletion, cells were co-transfected with 0.5 μg of 4xmts-mNeonGreen (AddGene; Plasmid #98876) and 10 nM of KIF5B siRNA#1 (ID s68782) or siRNA#2 (ID s68781) (Thermo Fisher Scientific). .. Silencer Select Negative Control (Thermo Fisher Scientific; #4390843) was used as the control. e3 Developmental Cell 60, 2962–2975.e1–e5, November 3, 2025

    Expressing:

    Article Title: The Role of the Tyrosine-Based Sorting Signals of the ORF3a Protein of SARS-CoV-2 in Intracellular Trafficking and Pathogenesis
    Article Snippet: .. To examine the co-localization of the ORF3a proteins with mitochondria, COS-7 cells were co-transfected with vectors expressing the ORF3a proteins and a vector expressing 4xmts-NeonGreen (mitochondria; Addgene, #98876). ..

    Article Title: The Role of the Tyrosine-Based Sorting Signals of the ORF3a Protein of SARS-CoV-2 in Intracellular Trafficking and Pathogenesis
    Article Snippet: .. To examine the co-localization of the ORF3a proteins with mitochondria, COS-7 cells were co-transfected with vectors expressing the ORF3a proteins and a vector expressing 4xmts-NeonGreen (mitochondria; Addgene, #98876). ..



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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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    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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    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