grx1-rogfp2 sensor Search Results


grx1 rogfp2 sensor  (Addgene inc)
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Addgene inc grx1 rogfp2 sensor
Oxidation induced by exogenous H 2 O 2 results in a redox change within the PV and parasite cytosol. (A) Schematic representation of the redox sensor based on the catalytic domain of glutaredoxin <t>(GRX1)</t> fused with ro-GFP2 depicting how this sensor interacts with GSH/GSSG and the excitation and emission values of the reduced and oxidized states. (B, C) Proof-of-principle tracking dynamic changes of GSH/GSSG during an oxidation event in <t>RH-GRX1-roGFP2</t> parasites. (B) Representative signal trace from the GRX1-roGFP2 sensor monitoring both fluorescence channels for reduced (red line) and oxidized (orange line) readouts over time following treatment with 10 mM H 2 O 2 . Microscopy images of an infected vacuole presented in pseudocolor (orange for oxidation, red for reduction) at a: baseline (45s), b: at the peak of the oxidation event (100s) and c: the return to baseline (280s). This is a representative trace from two independent experiments, with eight vacuoles. Scale bar: 5 µm. (C) Presents the oxidation/reduction ratio of normalized signal from graph (B) depicting the intensity changes in redox compared to baseline upon addition of 10 mM H 2 O 2 . (D) Representative trace from RH-GRA8-GRX1-roGFP2 parasites depicting 100 μM H 2 O 2 induced redox change within the PV. Microscopy images of an infected vacuole highlighting the presence of the sensor within PV (left image), alongside the brightfield image of the infected host cell. (E) A representative trace from RH-GRX1-roGFP2 parasites depicting 100 μM H 2 O 2 induction of redox change within the parasite cytosol. Microscopy images depicting an infected vacuole highlighting the presence of the sensor on cytosol (left panel image) alongside the brightfield image of the infected host cell. (D, E) Representative traces from three independent experiment, nine vacuoles for each group. Scale bar: 5 µm. (F, G) Representative trace of redox fluctuations in intracellular parasite following 1 μM ionomycin treatment for RH-GRA8-GRX1-roGFP2 (F) , and RH-GRX1-roGFP2 parasites (G) . (F, G) red arrows indicate the moment of parasite egress. Representative traces from three independent experiments, nine vacuoles for each group.
Grx1 Rogfp2 Sensor, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/grx1-rogfp2+sensor/pEIGW+Grx1-roGFP2+(Plasmid+%2364990)/pmc08382974-44-1-14
Average 93 stars, based on 1 article reviews
grx1 rogfp2 sensor - by Bioz Stars, 2026-09
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Oxidation induced by exogenous H 2 O 2 results in a redox change within the PV and parasite cytosol. (A) Schematic representation of the redox sensor based on the catalytic domain of glutaredoxin (GRX1) fused with ro-GFP2 depicting how this sensor interacts with GSH/GSSG and the excitation and emission values of the reduced and oxidized states. (B, C) Proof-of-principle tracking dynamic changes of GSH/GSSG during an oxidation event in RH-GRX1-roGFP2 parasites. (B) Representative signal trace from the GRX1-roGFP2 sensor monitoring both fluorescence channels for reduced (red line) and oxidized (orange line) readouts over time following treatment with 10 mM H 2 O 2 . Microscopy images of an infected vacuole presented in pseudocolor (orange for oxidation, red for reduction) at a: baseline (45s), b: at the peak of the oxidation event (100s) and c: the return to baseline (280s). This is a representative trace from two independent experiments, with eight vacuoles. Scale bar: 5 µm. (C) Presents the oxidation/reduction ratio of normalized signal from graph (B) depicting the intensity changes in redox compared to baseline upon addition of 10 mM H 2 O 2 . (D) Representative trace from RH-GRA8-GRX1-roGFP2 parasites depicting 100 μM H 2 O 2 induced redox change within the PV. Microscopy images of an infected vacuole highlighting the presence of the sensor within PV (left image), alongside the brightfield image of the infected host cell. (E) A representative trace from RH-GRX1-roGFP2 parasites depicting 100 μM H 2 O 2 induction of redox change within the parasite cytosol. Microscopy images depicting an infected vacuole highlighting the presence of the sensor on cytosol (left panel image) alongside the brightfield image of the infected host cell. (D, E) Representative traces from three independent experiment, nine vacuoles for each group. Scale bar: 5 µm. (F, G) Representative trace of redox fluctuations in intracellular parasite following 1 μM ionomycin treatment for RH-GRA8-GRX1-roGFP2 (F) , and RH-GRX1-roGFP2 parasites (G) . (F, G) red arrows indicate the moment of parasite egress. Representative traces from three independent experiments, nine vacuoles for each group.

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Extracellular Redox Signal Triggers Calcium Release and Impacts the Asexual Development of Toxoplasma gondii

doi: 10.3389/fcimb.2021.728425

Figure Lengend Snippet: Oxidation induced by exogenous H 2 O 2 results in a redox change within the PV and parasite cytosol. (A) Schematic representation of the redox sensor based on the catalytic domain of glutaredoxin (GRX1) fused with ro-GFP2 depicting how this sensor interacts with GSH/GSSG and the excitation and emission values of the reduced and oxidized states. (B, C) Proof-of-principle tracking dynamic changes of GSH/GSSG during an oxidation event in RH-GRX1-roGFP2 parasites. (B) Representative signal trace from the GRX1-roGFP2 sensor monitoring both fluorescence channels for reduced (red line) and oxidized (orange line) readouts over time following treatment with 10 mM H 2 O 2 . Microscopy images of an infected vacuole presented in pseudocolor (orange for oxidation, red for reduction) at a: baseline (45s), b: at the peak of the oxidation event (100s) and c: the return to baseline (280s). This is a representative trace from two independent experiments, with eight vacuoles. Scale bar: 5 µm. (C) Presents the oxidation/reduction ratio of normalized signal from graph (B) depicting the intensity changes in redox compared to baseline upon addition of 10 mM H 2 O 2 . (D) Representative trace from RH-GRA8-GRX1-roGFP2 parasites depicting 100 μM H 2 O 2 induced redox change within the PV. Microscopy images of an infected vacuole highlighting the presence of the sensor within PV (left image), alongside the brightfield image of the infected host cell. (E) A representative trace from RH-GRX1-roGFP2 parasites depicting 100 μM H 2 O 2 induction of redox change within the parasite cytosol. Microscopy images depicting an infected vacuole highlighting the presence of the sensor on cytosol (left panel image) alongside the brightfield image of the infected host cell. (D, E) Representative traces from three independent experiment, nine vacuoles for each group. Scale bar: 5 µm. (F, G) Representative trace of redox fluctuations in intracellular parasite following 1 μM ionomycin treatment for RH-GRA8-GRX1-roGFP2 (F) , and RH-GRX1-roGFP2 parasites (G) . (F, G) red arrows indicate the moment of parasite egress. Representative traces from three independent experiments, nine vacuoles for each group.

Article Snippet: The GRX1-roGFP2 sensor ( ; ) was amplified from the commercially available vector pEIGW-GRX1-roGFP2 (Addgene plasmid n°64990) using primers 1/2 and insert into digested (HF-EcoRI & PacI) pTUB8 vector containing selectable marker for HXGPRT using Gibson Assembly ® Master Mix.

Techniques: Fluorescence, Microscopy, Infection

The GRX1-roGFP redox sensor affects T gondii asexual replication. (A) Bar graphs presenting plaque count data from a six-day plaque assay using RH-GFP-Luc parasites as a reference control group. (B) Representative images of plaques formed. Small plaques are indicated by white arrows. Scale bar: 2 mm. (C) Violin plot presenting the distribution of plaque areas (mm 2 ) for parasites expressing the redox sensor within the cytosol. (D) Histogram presenting the effect of the GRX1-roGFP sensors and NAC on parasite intracellular replication. (A–D) All obtained from three independent experiments, with three technical replicates. Significance was calculated using one-way Anova, Bonferroni’s multiple comparisons test for (A, B) . P values: **< 0.01, **< 0.001 and ****< 0.0001. The significance analyse for (D) is provide on .

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: An Extracellular Redox Signal Triggers Calcium Release and Impacts the Asexual Development of Toxoplasma gondii

doi: 10.3389/fcimb.2021.728425

Figure Lengend Snippet: The GRX1-roGFP redox sensor affects T gondii asexual replication. (A) Bar graphs presenting plaque count data from a six-day plaque assay using RH-GFP-Luc parasites as a reference control group. (B) Representative images of plaques formed. Small plaques are indicated by white arrows. Scale bar: 2 mm. (C) Violin plot presenting the distribution of plaque areas (mm 2 ) for parasites expressing the redox sensor within the cytosol. (D) Histogram presenting the effect of the GRX1-roGFP sensors and NAC on parasite intracellular replication. (A–D) All obtained from three independent experiments, with three technical replicates. Significance was calculated using one-way Anova, Bonferroni’s multiple comparisons test for (A, B) . P values: **< 0.01, **< 0.001 and ****< 0.0001. The significance analyse for (D) is provide on .

Article Snippet: The GRX1-roGFP2 sensor ( ; ) was amplified from the commercially available vector pEIGW-GRX1-roGFP2 (Addgene plasmid n°64990) using primers 1/2 and insert into digested (HF-EcoRI & PacI) pTUB8 vector containing selectable marker for HXGPRT using Gibson Assembly ® Master Mix.

Techniques: Plaque Assay, Control, Expressing