phenazine methosulfate Search Results


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  • 99
    Millipore phenazine methosulfate
    The Δ soxR S. coelicolor mutant is not hypersensitive to peroxide or superoxide-generating agents. Paper disks soaked with solutions of 4-nitroquinoline, phenazine <t>methosulfate,</t> paraquat, or hydrogen peroxide were placed on bacterial lawns of wild-type
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    Promega phenazine methosulfate
    The Δ soxR S. coelicolor mutant is not hypersensitive to peroxide or superoxide-generating agents. Paper disks soaked with solutions of 4-nitroquinoline, phenazine <t>methosulfate,</t> paraquat, or hydrogen peroxide were placed on bacterial lawns of wild-type
    Phenazine Methosulfate, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega phenazine methosulfate pms
    Caspase-mediated cell death is not induced by tilapia piscidin (TP) 4. U87MG (wild-type p53 ) and U251 (mutant p53 ) cells were treated with different doses of TP4 (0, 5, 10, 15, 20, 50, and 100 μg/mL) for 24 h. Cell number ( A ) and cell viability ( B ) were assessed. Cell number was determined using the trypan exclusion assay. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt)/phenazine <t>methosulfate</t> <t>(MTS/PMS)</t> assay. Human umbilical vein endothelial cells (HUVECs) ( C ) and N27 rat neural precursor cells ( D ) were treated with indicated doses of TP4 for 24 h. Cell viability was assessed using MTS/PMS assay. U87MG and U251 cells were treated with TP4 (20 μg/mL) and staurosporine (1 μM) for 24 h and 6 h, respectively. Apoptosis was monitored by chromatin condensation ( E ), Annexin V binding ( F ), and caspase activation ( G ). Condensed nuclei were scored from 4′,6-diamidino-2-phenylindole (DAPI)-stained cells. Intensity of Annexin V-Fluorescein isothiocyanate (FITC) was measured by flow cytometry. Cell lysates were collected and immunoblotted with caspase-3, -8 and -9 antibodies. β-actin was used as a loading control. Right and bottom panel: ratio of cleaved caspase-3, -8, and -9 to procaspase-3, -8, and -9, respectively. Veh: vehicle; TP: TP4; Sta: staurosporine. ( H ) Cells were treated with TP4 (20 μg/mL) for different times. Cell lysates were immunoblotted with caspase-3 antibody. ( I ) Cells were preincubated with pan-caspase inhibitor Z-VAD fluoromethylketone (Z-VAD-FMK) (100 μM) for 1 h, followed by TP4 (20 μg/mL) and staurosporine treatment for 24 h and 6 h, respectively. Cell number was determined using the trypan exclusion assay. Vehicle: 0.5% dimethyl sulfoxide (DMSO). Western blotting experiments were performed at least three times with similar results. Band intensities were quantified with Image J software (1.51j8; NIH, Bethesda, MD, USA). * p
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    Thermo Fisher phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Thermo Fisher phenazine methosulfate pms
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Dojindo Labs 1 methoxy phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Millipore 1 methoxy 5 methylphenazinium methyl sulfate pms
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Promega electron coupling reagent phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Applichem 1 methoxy 5 methylphenazinium methyl sulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Promega 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
    3 4 5 Dimethylthiazol 2 Yl 2 5 Diphenyltetrazolium Bromide Phenazine Methosulfate, supplied by Promega, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore electron coupling reagent 5 methylphenazinium methyl sulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
    Electron Coupling Reagent 5 Methylphenazinium Methyl Sulfate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega phenazine methosulfate pms 20 1 solution
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Promega phenazine methosulfate pms solution
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
    Phenazine Methosulfate Pms Solution, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Welgene inc phenazine methosulfate pms reagent
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Fisher Scientific phenazine methosulfate pms
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    FUJIFILM phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
    Phenazine Methosulfate, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nacalai phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    TCI Tokyo Chemical Industry phenazine methosulfate
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
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    Promega inner salt phenazine methosulfate mts pms microtiter plate assay
    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine <t>methosulfate</t> (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P
    Inner Salt Phenazine Methosulfate Mts Pms Microtiter Plate Assay, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Δ soxR S. coelicolor mutant is not hypersensitive to peroxide or superoxide-generating agents. Paper disks soaked with solutions of 4-nitroquinoline, phenazine methosulfate, paraquat, or hydrogen peroxide were placed on bacterial lawns of wild-type

    Journal: Journal of Bacteriology

    Article Title: Expression of the Streptomyces coelicolor SoxR Regulon Is Intimately Linked with Actinorhodin Production ▿ SoxR Regulon Is Intimately Linked with Actinorhodin Production ▿ †

    doi: 10.1128/JB.00916-10

    Figure Lengend Snippet: The Δ soxR S. coelicolor mutant is not hypersensitive to peroxide or superoxide-generating agents. Paper disks soaked with solutions of 4-nitroquinoline, phenazine methosulfate, paraquat, or hydrogen peroxide were placed on bacterial lawns of wild-type

    Article Snippet: The drugs tested were paraquat, 4-nitroquinoline, phenazine methosulfate, plumbagin, menadione, diamide, H2 O2 , and tert -butyl hydroperoxide at the concentrations indicated (all reagents were purchased from Sigma).

    Techniques: Mutagenesis

    Caspase-mediated cell death is not induced by tilapia piscidin (TP) 4. U87MG (wild-type p53 ) and U251 (mutant p53 ) cells were treated with different doses of TP4 (0, 5, 10, 15, 20, 50, and 100 μg/mL) for 24 h. Cell number ( A ) and cell viability ( B ) were assessed. Cell number was determined using the trypan exclusion assay. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt)/phenazine methosulfate (MTS/PMS) assay. Human umbilical vein endothelial cells (HUVECs) ( C ) and N27 rat neural precursor cells ( D ) were treated with indicated doses of TP4 for 24 h. Cell viability was assessed using MTS/PMS assay. U87MG and U251 cells were treated with TP4 (20 μg/mL) and staurosporine (1 μM) for 24 h and 6 h, respectively. Apoptosis was monitored by chromatin condensation ( E ), Annexin V binding ( F ), and caspase activation ( G ). Condensed nuclei were scored from 4′,6-diamidino-2-phenylindole (DAPI)-stained cells. Intensity of Annexin V-Fluorescein isothiocyanate (FITC) was measured by flow cytometry. Cell lysates were collected and immunoblotted with caspase-3, -8 and -9 antibodies. β-actin was used as a loading control. Right and bottom panel: ratio of cleaved caspase-3, -8, and -9 to procaspase-3, -8, and -9, respectively. Veh: vehicle; TP: TP4; Sta: staurosporine. ( H ) Cells were treated with TP4 (20 μg/mL) for different times. Cell lysates were immunoblotted with caspase-3 antibody. ( I ) Cells were preincubated with pan-caspase inhibitor Z-VAD fluoromethylketone (Z-VAD-FMK) (100 μM) for 1 h, followed by TP4 (20 μg/mL) and staurosporine treatment for 24 h and 6 h, respectively. Cell number was determined using the trypan exclusion assay. Vehicle: 0.5% dimethyl sulfoxide (DMSO). Western blotting experiments were performed at least three times with similar results. Band intensities were quantified with Image J software (1.51j8; NIH, Bethesda, MD, USA). * p

    Journal: Cancers

    Article Title: Antimicrobial Peptide TP4 Induces ROS-Mediated Necrosis by Triggering Mitochondrial Dysfunction in Wild-Type and Mutant p53 Glioblastoma Cells

    doi: 10.3390/cancers11020171

    Figure Lengend Snippet: Caspase-mediated cell death is not induced by tilapia piscidin (TP) 4. U87MG (wild-type p53 ) and U251 (mutant p53 ) cells were treated with different doses of TP4 (0, 5, 10, 15, 20, 50, and 100 μg/mL) for 24 h. Cell number ( A ) and cell viability ( B ) were assessed. Cell number was determined using the trypan exclusion assay. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt)/phenazine methosulfate (MTS/PMS) assay. Human umbilical vein endothelial cells (HUVECs) ( C ) and N27 rat neural precursor cells ( D ) were treated with indicated doses of TP4 for 24 h. Cell viability was assessed using MTS/PMS assay. U87MG and U251 cells were treated with TP4 (20 μg/mL) and staurosporine (1 μM) for 24 h and 6 h, respectively. Apoptosis was monitored by chromatin condensation ( E ), Annexin V binding ( F ), and caspase activation ( G ). Condensed nuclei were scored from 4′,6-diamidino-2-phenylindole (DAPI)-stained cells. Intensity of Annexin V-Fluorescein isothiocyanate (FITC) was measured by flow cytometry. Cell lysates were collected and immunoblotted with caspase-3, -8 and -9 antibodies. β-actin was used as a loading control. Right and bottom panel: ratio of cleaved caspase-3, -8, and -9 to procaspase-3, -8, and -9, respectively. Veh: vehicle; TP: TP4; Sta: staurosporine. ( H ) Cells were treated with TP4 (20 μg/mL) for different times. Cell lysates were immunoblotted with caspase-3 antibody. ( I ) Cells were preincubated with pan-caspase inhibitor Z-VAD fluoromethylketone (Z-VAD-FMK) (100 μM) for 1 h, followed by TP4 (20 μg/mL) and staurosporine treatment for 24 h and 6 h, respectively. Cell number was determined using the trypan exclusion assay. Vehicle: 0.5% dimethyl sulfoxide (DMSO). Western blotting experiments were performed at least three times with similar results. Band intensities were quantified with Image J software (1.51j8; NIH, Bethesda, MD, USA). * p

    Article Snippet: 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) and phenazine methosulfate (PMS) were purchased from Promega (Madison, WI, USA).

    Techniques: Mutagenesis, Exclusion Assay, Binding Assay, Activation Assay, Staining, Flow Cytometry, Cytometry, Western Blot, Software

    C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine methosulfate (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P

    Journal: mSphere

    Article Title: Bacteria Modify Candida albicans Hypha Formation, Microcolony Properties, and Survival within Macrophages

    doi: 10.1128/mSphere.00689-20

    Figure Lengend Snippet: C. albicans microcolony density is decreased by P. aeruginosa and phenazines. (A) C. albicans microcolonies were cultured using RPMI 1640 medium at 37°C in 5% CO 2 , along with P. aeruginosa strains POA1, 0635, or PA14 or of fresh-filtered P. aeruginosa culture supernatant (10%; s Pa ), phenazine methosulfate (PMS; 5 μM), or pyocyanin (PYO; 20 μM) for 17 h. C. albicans incubated alone was used as control. Images were obtained using bright-field microscopy, and the microcolony density was calculated using ImageJ. (B) The microcolony density per square micron was obtained by using ImageJ. The addition of three P. aeruginosa strains or s Pa each significantly decreased microcolony density compared to C. albicans grown in culture media alone. Experiments were carried out in duplicate, and the means ± the SD of four independent experiments are shown. Significance was obtained by Student t test or one-way ANOVA with post ad hoc Dunnett’s multiple-comparison test (*, P

    Article Snippet: For experiments with phenazines, purified pyocyanin (30 mM) (PYO; Cayman Chemical) and phenazine methosulfate (25 mM; PMS; Acros Organic) were diluted in H2 O to reach a final concentration after addition to wells (PYO [20 μM] and PMS [5 μM]).

    Techniques: Cell Culture, Incubation, Microscopy