kt5720  (Tocris)

Journal: Scientific Reports

Article Title: Tectorigenin induces vasorelaxation in porcine coronary arteries through activation of Kv channels and oestrogen receptor modulation

doi: 10.1038/s41598-025-20988-6

Figure Lengend Snippet: Effects of neural and cyclic nucleotide pathway inhibitors on tectorigenin-induced relaxation in porcine coronary arteries pre-contracted with 100 nM U46619. ( A ) Tetrodotoxin (TTX, 1 µM) and ω-conotoxin GVIA (CTX, 1 µM) had no significant effect on the relaxation induced by 30 µM tectorigenin ( p > 0.05, n = 4). ( B ) Pretreatment with rolipram (1 µM, a selective phosphodiesterase-4 inhibitor) or vardenafil (1 µM, a selective phosphodiesterase-5 inhibitor) did not significantly alter tectorigenin-induced vasorelaxation ( p > 0.05, n = 4). ( C ) Inhibitors of the nitric oxide and cyclic nucleotide pathways, including Nω-nitro-L-arginine (L-NNA, 100 µM), KT5720 (1 µM, a PKA inhibitor), and KT5823 (1 µM, a PKG inhibitor), also did not significantly affect the relaxant response to tectorigenin ( p > 0.05, n = 4). Data are expressed as mean ± standard error of the mean (SEM) from four independent hearts. U46619 plateau (normalised to 60 mM KCl) was similar across groups ( p > 0.05; Supplementary Table 2).

Article Snippet: For experimental assays, a range of pharmacological agents was utilised, including U46619, apamin, KT5720, KT5823, and L-NNA (Sigma-Aldrich, MO, USA); rolipram, vardenafil, and TEA (Santa Cruz Biotechnology, CA, USA); IbTX (Alomone Labs, Jerusalem, Israel); glibenclamide (Research Biochemicals International, MA, USA); TTX and 4-AP (Tocris Bioscience, Bristol, UK); CTX (Bachem, Bubendorf, Switzerland); and charybdotoxin, methyl-piperidino-pyrazole (MPP), and PHTPP (Cayman Chemical, MI, USA).

Techniques:

Journal: PLOS ONE

Article Title: Role of canonical and non-canonical cAMP sources in CRHR2α-dependent signaling

doi: 10.1371/journal.pone.0310699

Figure Lengend Snippet: Cells were stimulated with 100 nM UCN1 (blue) or UCN3 (green) for the indicated time points in the presence or absence of a, c, e PKA activity inhibitor (10 μM H89) or b, d, f MEK inhibitor (10 μM U0126) a-d , CREB and Actin were measured by immunoblotting and quantified by densitometry using Fiji ImageJ software. pCREB was normalized to Actin. Results are expressed as the percentage of maximum pCREB obtained after stimulation. Data: mean± SEM, 3 independent experiments, *p<0,05, **p<0,01 with respect to basal by two ways ANOVA following by Tukey test. e, f, c-fos mRNA levels after pre-incubation with inhibitors (10 μM H89 or U0126) and 45 minutes of stimulation with 100 nM UCN1 or UCN3 were determined by RT PCR and normalized to Hprt. Data: Mean ± SEM, n = 3, ***p<0,001 with respect to basal by one way ANOVA following by tukey test.

Article Snippet: The following inhibitors were used: H89 and KT5720 (PKA; 371963, Calbiochem, HY-N6789, MedChem Express respectively), 2′,5′-dideoxyadenosine (tmACs; 288104, Calbiochem), KH7 and LRE1 (sAC; 3834, Tocris, HY-100524 MedChem Express respectively), U0126 (MEK1/2; 662005, Calbiochem).

Techniques: Activity Assay, Western Blot, Software, Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: PLOS ONE

Article Title: Role of canonical and non-canonical cAMP sources in CRHR2α-dependent signaling

doi: 10.1371/journal.pone.0310699

Figure Lengend Snippet: Cells were stimulated with 100 nM UCN1 (blue) or UCN3 (green) in presence or absence of a, b , PKA activity inhibitor (10 μM H89) or c, d, MEK inhibitor (10 μM U0126). a, c , Neurite outgrowth was determined per cell after 15 min-pretreatment with inhibitors and 1h-treatment with the agonists, as the ratio between the longest neurite and the soma in each cell. Data: mean ± SEM, n = 3. ***, p<0,001 ns: no significative with respect to basal by repeated measures one-way ANOVA following by Tukey test. b, d , Representative photographs are shown for each treatment. Bars 20 μM.

Article Snippet: The following inhibitors were used: H89 and KT5720 (PKA; 371963, Calbiochem, HY-N6789, MedChem Express respectively), 2′,5′-dideoxyadenosine (tmACs; 288104, Calbiochem), KH7 and LRE1 (sAC; 3834, Tocris, HY-100524 MedChem Express respectively), U0126 (MEK1/2; 662005, Calbiochem).

Techniques: Activity Assay