ns 2028  (Tocris)

Journal: Journal of cell communication and signaling

Article Title: Mutant p53 gain-of-function stimulates canonical Wnt signaling via PI3K/AKT pathway in colon cancer.

doi: 10.1007/s12079-023-00793-4

Figure Lengend Snippet: Fig. 3 Mutant p53 knockdown in the SW480 cell line decreases canonical Wnt pathway. A Comparation of non-phospho- rylated β-catenin (active) and p53 protein levels measured in different colon cancer cell lines with different TP53 status, RKO (wild-type p53), SW480 (R273H p53), and SW620 (R273H p53). Densitometric analysis of immunoblots of p53 and active β-catenin was performed using GAPDH as a loading control. B Representa- tive immunoblots and graph of p53 protein in SW480 under different doses of siRNA p53 (25–100 nM); the scramble condition was used as a control (100 nM). C Luciferase activity measured with the TOP/FOP system in both scramble and siRNA p53 (100 nM) condi- tions. The relative units were normalized to Renilla activity. D Representative immunoblot and graph of c-myc normalized to GAPDH. E. Representa- tive colonies and relative area quantification corresponding to scramble and shp53 conditions of SW480-transduced cells. Graphs represent densito- metric analysis from at least three independent experiments (means ± SEM). *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The PI3K/AKT inhibitor wortmannin was obtained from Sigma (Cat. No. 19545-26-7), and the p53 Activator III compound RITA was from Santa Cruz Biotechnology (Cat. No. sc-202743).

Techniques: Mutagenesis, Knockdown, Western Blot, Control, Luciferase, Activity Assay

Journal: PLoS ONE

Article Title: Fluorescence Dequenching Makes Haem-Free Soluble Guanylate Cyclase Detectable in Living Cells

doi: 10.1371/journal.pone.0023596

Figure Lengend Snippet: cGMP reporter cells were cotransfected with WT α 1 sGC and TC4-WT β 1 sGC and labelled with FlAsH. The cells were then incubated with 0.1, 1 or 10 µM BAY 58-2667 (A) 1, 30 or 100 µM rotenone (B) or 1, 10 or 100 µM NS 2028 (C) for 90 min. Fluorescence intensity of single cells was monitored in a time series of 120 laser scans and is expressed as % of start value which was obtained before application of the test compounds. Data are means ± S.E.M. from 9–28 single cells assayed on different days. * p <0.05; *** p <0.005: Student's t -test compared to control. Representative traces of the fluorescence measurement as % of start value are given in the right part of the figure.

Article Snippet: ODQ (1H-(1,2,4)-oxadiazole(4,3- a )quinoxalin-1-one) was purchased from Tocris Bioscience (Avonmouth, UK) and NS 2028 (4 H -8-Bromo-1,2,4-oxadiazolo(3,4-d)benz(b)(1,4)oxazin-1-one) was purchased from Axxora GmbH (Loerrach, Germany).

Techniques: Incubation, Fluorescence

Journal: PLoS ONE

Article Title: Fluorescence Dequenching Makes Haem-Free Soluble Guanylate Cyclase Detectable in Living Cells

doi: 10.1371/journal.pone.0023596

Figure Lengend Snippet: Effects of 100 µM NS 2028 on fluorescence intensity of FlAsH labelled haem- free TC4-Y135A/R139A sGC (A) and ReAsH labelled TC4-WT sGC (B) after 90 min incubation.

Article Snippet: ODQ (1H-(1,2,4)-oxadiazole(4,3- a )quinoxalin-1-one) was purchased from Tocris Bioscience (Avonmouth, UK) and NS 2028 (4 H -8-Bromo-1,2,4-oxadiazolo(3,4-d)benz(b)(1,4)oxazin-1-one) was purchased from Axxora GmbH (Loerrach, Germany).

Techniques: Fluorescence, Incubation

Journal: PLoS ONE

Article Title: Fluorescence Dequenching Makes Haem-Free Soluble Guanylate Cyclase Detectable in Living Cells

doi: 10.1371/journal.pone.0023596

Figure Lengend Snippet: cGMP reporter cells expressing TC4-WT sGC were pre-incubated with 100 µM NS 2028 or rotenone and then incubated with increasing concentrations of BAY 58-2667, BAY 41-2272 alone or in combination with 10 µM ODQ or 10 nM DEA/NO (NO), respectively. Enzyme activity is expressed as x-fold stimulation compared to pre-treated but not stimulated control. Data are means ± S.E.M. from 3–10 independent experiments, performed in duplicate. A basal activity of 7747 relative light units was measured. A) is identical to .

Article Snippet: ODQ (1H-(1,2,4)-oxadiazole(4,3- a )quinoxalin-1-one) was purchased from Tocris Bioscience (Avonmouth, UK) and NS 2028 (4 H -8-Bromo-1,2,4-oxadiazolo(3,4-d)benz(b)(1,4)oxazin-1-one) was purchased from Axxora GmbH (Loerrach, Germany).

Techniques: Expressing, Incubation, Activity Assay

Journal: PLoS ONE

Article Title: Fluorescence Dequenching Makes Haem-Free Soluble Guanylate Cyclase Detectable in Living Cells

doi: 10.1371/journal.pone.0023596

Figure Lengend Snippet: Effects of 90 min pre-treatment with 100 µM NS 2028 or rotenone on TC4-WT sGC protein levels.

Article Snippet: ODQ (1H-(1,2,4)-oxadiazole(4,3- a )quinoxalin-1-one) was purchased from Tocris Bioscience (Avonmouth, UK) and NS 2028 (4 H -8-Bromo-1,2,4-oxadiazolo(3,4-d)benz(b)(1,4)oxazin-1-one) was purchased from Axxora GmbH (Loerrach, Germany).

Techniques: