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    MedChemExpress cinobufagin
    Effect of <t>cinobufagin</t> on ANO1 chloride channel activity and intestinal smooth muscle contraction. ( A ) Representative apical membrane currents from 3 independent experiments in FRT cells expressing ANO1. Indicated concentrations of cinobufagin and 10 μM Ani9-5f were pretreated for 10 min. ANO1 chloride channel was activated by 100 μM ATP. ( B ) Representative smooth muscle contraction traces from 4 independent experiments with mouse ileal segments. Indicated concentrations of cinobufagin and 10 μM Ani9-5f applied to mouse ileal segments.
    Cinobufagin, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Effect of cinobufagin on ANO1 chloride channel activity and intestinal smooth muscle contraction. ( A ) Representative apical membrane currents from 3 independent experiments in FRT cells expressing ANO1. Indicated concentrations of cinobufagin and 10 μM Ani9-5f were pretreated for 10 min. ANO1 chloride channel was activated by 100 μM ATP. ( B ) Representative smooth muscle contraction traces from 4 independent experiments with mouse ileal segments. Indicated concentrations of cinobufagin and 10 μM Ani9-5f applied to mouse ileal segments.

    Journal: International Journal of Molecular Sciences

    Article Title: Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1

    doi: 10.3390/ijms222112037

    Figure Lengend Snippet: Effect of cinobufagin on ANO1 chloride channel activity and intestinal smooth muscle contraction. ( A ) Representative apical membrane currents from 3 independent experiments in FRT cells expressing ANO1. Indicated concentrations of cinobufagin and 10 μM Ani9-5f were pretreated for 10 min. ANO1 chloride channel was activated by 100 μM ATP. ( B ) Representative smooth muscle contraction traces from 4 independent experiments with mouse ileal segments. Indicated concentrations of cinobufagin and 10 μM Ani9-5f applied to mouse ileal segments.

    Article Snippet: Cinobufagin was purchased from MedChem Express (Monmouth Junction, NJ, USA).

    Techniques: Activity Assay, Expressing

    Effect of cinobufagin on protein expression level of ANO1 and CFTR. ( A ) Representative immunoblot analysis of ANO1 in cinobufagin-treated CAL-27 cells. Cinobufagin-treated cells at the indicated concentration for 24 h. CRISPR/Cas9 technique was used to generate CAL-27 ANO1 knockout (KO) cells. ( B ) ANO1 protein intensities were normalized to those of β-actin (mean ± S.D., n = 4). * p

    Journal: International Journal of Molecular Sciences

    Article Title: Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1

    doi: 10.3390/ijms222112037

    Figure Lengend Snippet: Effect of cinobufagin on protein expression level of ANO1 and CFTR. ( A ) Representative immunoblot analysis of ANO1 in cinobufagin-treated CAL-27 cells. Cinobufagin-treated cells at the indicated concentration for 24 h. CRISPR/Cas9 technique was used to generate CAL-27 ANO1 knockout (KO) cells. ( B ) ANO1 protein intensities were normalized to those of β-actin (mean ± S.D., n = 4). * p

    Article Snippet: Cinobufagin was purchased from MedChem Express (Monmouth Junction, NJ, USA).

    Techniques: Expressing, Concentration Assay, CRISPR, Knock-Out

    Identification of novel compounds that downregulate ANO in CAL-27 cells. ( A ) Principle of high-throughput screening assay. ( B ) Representative YFP fluorescence traces. YFP fluorescence monitored in CAL-27 cells treated with 20 μM test compounds for 24 h. (right) Dot plot of primary screening results for 730 natural products. ( C ) Representative YFP fluorescence traces showing the effect of short-term (10 min) exposure of 20 μM cinobufagin, fangchinoline and lanatoside C on ANO1 channel activity in CAL-27 cells. ( D ) Chemical structures of cinobufagin, fangchinoline and lanatoside C. ( E ) Effects of Ani9-5f, cinobufagin, fangchinoline and lanatoside C on ANO1 protein expression levels were assessed by Western blot analysis. CAL-27 cells were treated with 10 μM of the test compounds for 24 h.

    Journal: International Journal of Molecular Sciences

    Article Title: Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1

    doi: 10.3390/ijms222112037

    Figure Lengend Snippet: Identification of novel compounds that downregulate ANO in CAL-27 cells. ( A ) Principle of high-throughput screening assay. ( B ) Representative YFP fluorescence traces. YFP fluorescence monitored in CAL-27 cells treated with 20 μM test compounds for 24 h. (right) Dot plot of primary screening results for 730 natural products. ( C ) Representative YFP fluorescence traces showing the effect of short-term (10 min) exposure of 20 μM cinobufagin, fangchinoline and lanatoside C on ANO1 channel activity in CAL-27 cells. ( D ) Chemical structures of cinobufagin, fangchinoline and lanatoside C. ( E ) Effects of Ani9-5f, cinobufagin, fangchinoline and lanatoside C on ANO1 protein expression levels were assessed by Western blot analysis. CAL-27 cells were treated with 10 μM of the test compounds for 24 h.

    Article Snippet: Cinobufagin was purchased from MedChem Express (Monmouth Junction, NJ, USA).

    Techniques: High Throughput Screening Assay, Screening Assay, Fluorescence, Activity Assay, Expressing, Western Blot

    Effect of cinobufagin on caspase-3 activation and induction of PARP cleavage in CAL-27 cells. ( A ) Images were taken 24 h after application of 300 nM cinobufagin. Cells were incubated with caspase-3 substrate (green) and Hoechst 33342 (blue) 20 min prior to image acquisition. Scale bars represent 200 μm. ( B ) CAL-27 cells were treated with the indicated concentrations of cinobufagin in the presence or absence of 10 μM of Ac-DEVD-CHO for 24 h, then the cells were treated with 2 μM of caspase-3 substrate for 20 min to estimate caspase-3 activity (mean ± S.E., n = 3). ( C ) Representative immunoblot analysis of PARP and cleaved PARP(C-PARP). Cells were treated with the indicated concentrations of cinobufagin for 24 h. ( D ) Cleaved PARP protein intensities were normalized to those of β-actin (mean ± S.D., n = 3). * p

    Journal: International Journal of Molecular Sciences

    Article Title: Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1

    doi: 10.3390/ijms222112037

    Figure Lengend Snippet: Effect of cinobufagin on caspase-3 activation and induction of PARP cleavage in CAL-27 cells. ( A ) Images were taken 24 h after application of 300 nM cinobufagin. Cells were incubated with caspase-3 substrate (green) and Hoechst 33342 (blue) 20 min prior to image acquisition. Scale bars represent 200 μm. ( B ) CAL-27 cells were treated with the indicated concentrations of cinobufagin in the presence or absence of 10 μM of Ac-DEVD-CHO for 24 h, then the cells were treated with 2 μM of caspase-3 substrate for 20 min to estimate caspase-3 activity (mean ± S.E., n = 3). ( C ) Representative immunoblot analysis of PARP and cleaved PARP(C-PARP). Cells were treated with the indicated concentrations of cinobufagin for 24 h. ( D ) Cleaved PARP protein intensities were normalized to those of β-actin (mean ± S.D., n = 3). * p

    Article Snippet: Cinobufagin was purchased from MedChem Express (Monmouth Junction, NJ, USA).

    Techniques: Activation Assay, Incubation, Activity Assay

    Effects of cinobufagin on mRNA expression level of ANO1 and phosphorylation of STAT3 in CAL-27 cells. ( A ) mRNA expression levels of ANO1 were measured using real-time PCR in CAL-27 cells. Indicated concentrations of cinobufagin were treated for 24 h (mean ± S.E., n = 3). ( B ) Representative immunoblot analysis of pSTAT3 in CAL-27 cells treated with cinobufagin for 24 h. ( C ) Phosphorylated STAT3 (pSTAT3) levels were normalized against total STAT3 levels (mean ± S.E., n = 3). ( D ) Representative immunoblot analysis of pSTAT3 in CAL-27 cells treated with niclosamide at indicated concentrations for 24 h. ( E ) pSTAT3 levels were normalized against total STAT3 levels (mean ± S.E., n = 3). ( F ) mRNA expression levels of ANO1 were measured in CAL-27 cells treated with niclosamide at indicated concentrations for 24 h (mean ± S.E., n = 3). * p

    Journal: International Journal of Molecular Sciences

    Article Title: Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1

    doi: 10.3390/ijms222112037

    Figure Lengend Snippet: Effects of cinobufagin on mRNA expression level of ANO1 and phosphorylation of STAT3 in CAL-27 cells. ( A ) mRNA expression levels of ANO1 were measured using real-time PCR in CAL-27 cells. Indicated concentrations of cinobufagin were treated for 24 h (mean ± S.E., n = 3). ( B ) Representative immunoblot analysis of pSTAT3 in CAL-27 cells treated with cinobufagin for 24 h. ( C ) Phosphorylated STAT3 (pSTAT3) levels were normalized against total STAT3 levels (mean ± S.E., n = 3). ( D ) Representative immunoblot analysis of pSTAT3 in CAL-27 cells treated with niclosamide at indicated concentrations for 24 h. ( E ) pSTAT3 levels were normalized against total STAT3 levels (mean ± S.E., n = 3). ( F ) mRNA expression levels of ANO1 were measured in CAL-27 cells treated with niclosamide at indicated concentrations for 24 h (mean ± S.E., n = 3). * p

    Article Snippet: Cinobufagin was purchased from MedChem Express (Monmouth Junction, NJ, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Effect of cinobufagin on cell viability and migration in CAL-27 cells. ( A ) Effect of cinobufagin on cell viability in CAL-27, and ANO1 knockout (KO) CAL-27 cells. These cells were treated with cinobufagin at the indicated concentrations for 48 h, and MTS assay was performed to estimate cell viability (mean ± S.E., n = 5). ( B , C ) Scratch wound healing assay was conducted in CAL-27 cells expressing high levels of ANO1. Cells were treated with cinobufagin at the indicated concentration, and time-lapse images were acquired every 2 h after wound generation (mean ± S.E., n = 3). Scale bars represent 300 μm. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Cinobufagin Exerts Anticancer Activity in Oral Squamous Cell Carcinoma Cells through Downregulation of ANO1

    doi: 10.3390/ijms222112037

    Figure Lengend Snippet: Effect of cinobufagin on cell viability and migration in CAL-27 cells. ( A ) Effect of cinobufagin on cell viability in CAL-27, and ANO1 knockout (KO) CAL-27 cells. These cells were treated with cinobufagin at the indicated concentrations for 48 h, and MTS assay was performed to estimate cell viability (mean ± S.E., n = 5). ( B , C ) Scratch wound healing assay was conducted in CAL-27 cells expressing high levels of ANO1. Cells were treated with cinobufagin at the indicated concentration, and time-lapse images were acquired every 2 h after wound generation (mean ± S.E., n = 3). Scale bars represent 300 μm. * p

    Article Snippet: Cinobufagin was purchased from MedChem Express (Monmouth Junction, NJ, USA).

    Techniques: Migration, Knock-Out, MTS Assay, Wound Healing Assay, Expressing, Concentration Assay