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ml171  (MedChemExpress)
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MedChemExpress ml171
Effect of CTX1 on [Ca 2+ ]i and ROS level in U937 cells. U937 cells were treated with 500 nM CTX1 for 2 h (for measuring [Ca 2+ ]i) or 4 h (for measuring ROS level and cell viability). On the other hand, U937 cells were pretreated with 2 mM N-acetylcysteine (NAC), 10 μM BAPTA-AM, 10 μM GLX351322, 10 μM <t>ML171,</t> 10 μM EGTA, 10 μM ruthenium red (RR), or 50 μM 2-aminoethoxydiphenyl borane (2-APB) for 1 h and then incubated with 500 nM CTX1 for 2 h (for measuring [Ca 2+ ]i) or for 4 h (for measuring ROS level and cell viability). ( A ) CTX1 induced ROS generation in U937 cells. U937 cells were treated with 500 nM CTX1 for indicated time periods. Results were shown as fold-increase in fluorescence intensity compared with the control group. Each value is the mean ± SD of three independent experiments with triplicate measurements. ( B ) CTX1 induced an elevation of [Ca 2+ ]i in U937 cells. U937 cells were treated with 500 nM CTX1 for indicated time periods. Results were shown as fold-increase in fluorescence intensity compared with the control group. ( C ) Effect of BAPTA−AM, NAC, ML171, and GLX351322 on CTX1-induced ROS generation (mean ± SD, * p < 0.05; NS, not statistically significant). ( D ) Effect of BAPTA−AM, NAC, and GLX351322 on CTX1-induced an increase in [Ca 2+ ]i (mean ± SD, * p < 0.05; NS, not statistically significant). ( E ) Effect of EGTA, ruthenium red, and 2−APB on CTX1-induced an increase in [Ca 2+ ]i (mean ± SD, * p < 0.05; NS, not statistically significant). ( F ) Effect of BAPTA−AM, GLX351322, and NAC on the viability of CTX1-treated cells (mean ± SD, * p < 0.05). ( G ) Effect of CTX1 on NOX4 protein expression (* p < 0.05, CTX1-treated cells compared to untreated control cells). ( H ) Detecting the expression of NOX4 mRNA using qRT-PCR (mean ± SD, * p < 0.05). ( I ) Effect of CTX1 on the luciferase activity of NOX4 promoter construct. After transfection with indicated plasmid for 24 h, transfected cells were treated with 500 nM CTX1 for 4 h and then harvested for measuring luciferase activity (mean ± SD, * p < 0.05). ( J ) Effect of BAPTA−AM on the expression of NOX4 mRNA in CTX1-treated cells (mean ± SD, * p < 0.05). ( K ) Effect of BAPTA−AM on the expression of NOX4 protein in CTX1-treated cells (* p < 0.05, BAPTA−AM/CTX1-treated cells compared to CTX1-treated cells).
Ml171, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Effect of CTX1 on [Ca 2+ ]i and ROS level in U937 cells. U937 cells were treated with 500 nM CTX1 for 2 h (for measuring [Ca 2+ ]i) or 4 h (for measuring ROS level and cell viability). On the other hand, U937 cells were pretreated with 2 mM N-acetylcysteine (NAC), 10 μM BAPTA-AM, 10 μM GLX351322, 10 μM ML171, 10 μM EGTA, 10 μM ruthenium red (RR), or 50 μM 2-aminoethoxydiphenyl borane (2-APB) for 1 h and then incubated with 500 nM CTX1 for 2 h (for measuring [Ca 2+ ]i) or for 4 h (for measuring ROS level and cell viability). ( A ) CTX1 induced ROS generation in U937 cells. U937 cells were treated with 500 nM CTX1 for indicated time periods. Results were shown as fold-increase in fluorescence intensity compared with the control group. Each value is the mean ± SD of three independent experiments with triplicate measurements. ( B ) CTX1 induced an elevation of [Ca 2+ ]i in U937 cells. U937 cells were treated with 500 nM CTX1 for indicated time periods. Results were shown as fold-increase in fluorescence intensity compared with the control group. ( C ) Effect of BAPTA−AM, NAC, ML171, and GLX351322 on CTX1-induced ROS generation (mean ± SD, * p < 0.05; NS, not statistically significant). ( D ) Effect of BAPTA−AM, NAC, and GLX351322 on CTX1-induced an increase in [Ca 2+ ]i (mean ± SD, * p < 0.05; NS, not statistically significant). ( E ) Effect of EGTA, ruthenium red, and 2−APB on CTX1-induced an increase in [Ca 2+ ]i (mean ± SD, * p < 0.05; NS, not statistically significant). ( F ) Effect of BAPTA−AM, GLX351322, and NAC on the viability of CTX1-treated cells (mean ± SD, * p < 0.05). ( G ) Effect of CTX1 on NOX4 protein expression (* p < 0.05, CTX1-treated cells compared to untreated control cells). ( H ) Detecting the expression of NOX4 mRNA using qRT-PCR (mean ± SD, * p < 0.05). ( I ) Effect of CTX1 on the luciferase activity of NOX4 promoter construct. After transfection with indicated plasmid for 24 h, transfected cells were treated with 500 nM CTX1 for 4 h and then harvested for measuring luciferase activity (mean ± SD, * p < 0.05). ( J ) Effect of BAPTA−AM on the expression of NOX4 mRNA in CTX1-treated cells (mean ± SD, * p < 0.05). ( K ) Effect of BAPTA−AM on the expression of NOX4 protein in CTX1-treated cells (* p < 0.05, BAPTA−AM/CTX1-treated cells compared to CTX1-treated cells).

Journal: Cells

Article Title: Naja atra Cardiotoxin 1 Induces the FasL/Fas Death Pathway in Human Leukemia Cells

doi: 10.3390/cells10082073

Figure Lengend Snippet: Effect of CTX1 on [Ca 2+ ]i and ROS level in U937 cells. U937 cells were treated with 500 nM CTX1 for 2 h (for measuring [Ca 2+ ]i) or 4 h (for measuring ROS level and cell viability). On the other hand, U937 cells were pretreated with 2 mM N-acetylcysteine (NAC), 10 μM BAPTA-AM, 10 μM GLX351322, 10 μM ML171, 10 μM EGTA, 10 μM ruthenium red (RR), or 50 μM 2-aminoethoxydiphenyl borane (2-APB) for 1 h and then incubated with 500 nM CTX1 for 2 h (for measuring [Ca 2+ ]i) or for 4 h (for measuring ROS level and cell viability). ( A ) CTX1 induced ROS generation in U937 cells. U937 cells were treated with 500 nM CTX1 for indicated time periods. Results were shown as fold-increase in fluorescence intensity compared with the control group. Each value is the mean ± SD of three independent experiments with triplicate measurements. ( B ) CTX1 induced an elevation of [Ca 2+ ]i in U937 cells. U937 cells were treated with 500 nM CTX1 for indicated time periods. Results were shown as fold-increase in fluorescence intensity compared with the control group. ( C ) Effect of BAPTA−AM, NAC, ML171, and GLX351322 on CTX1-induced ROS generation (mean ± SD, * p < 0.05; NS, not statistically significant). ( D ) Effect of BAPTA−AM, NAC, and GLX351322 on CTX1-induced an increase in [Ca 2+ ]i (mean ± SD, * p < 0.05; NS, not statistically significant). ( E ) Effect of EGTA, ruthenium red, and 2−APB on CTX1-induced an increase in [Ca 2+ ]i (mean ± SD, * p < 0.05; NS, not statistically significant). ( F ) Effect of BAPTA−AM, GLX351322, and NAC on the viability of CTX1-treated cells (mean ± SD, * p < 0.05). ( G ) Effect of CTX1 on NOX4 protein expression (* p < 0.05, CTX1-treated cells compared to untreated control cells). ( H ) Detecting the expression of NOX4 mRNA using qRT-PCR (mean ± SD, * p < 0.05). ( I ) Effect of CTX1 on the luciferase activity of NOX4 promoter construct. After transfection with indicated plasmid for 24 h, transfected cells were treated with 500 nM CTX1 for 4 h and then harvested for measuring luciferase activity (mean ± SD, * p < 0.05). ( J ) Effect of BAPTA−AM on the expression of NOX4 mRNA in CTX1-treated cells (mean ± SD, * p < 0.05). ( K ) Effect of BAPTA−AM on the expression of NOX4 protein in CTX1-treated cells (* p < 0.05, BAPTA−AM/CTX1-treated cells compared to CTX1-treated cells).

Article Snippet: Without specific indication, the reagents used in this study were obtained from Sigma-Aldrich (St. Louis, MO, USA), and ML171 and GLX351322 were from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Incubation, Fluorescence, Control, Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, Construct, Transfection, Plasmid Preparation