Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cysteinyl-leukotriene receptor type 1 expression and function is down-regulated during monocyte-derived dendritic cell maturation with zymosan: involvement of IL-10 and prostaglandins.

doi: 10.4049/jimmunol.0901800

Figure Lengend Snippet: FIGURE 5. Involvement of the ERK and p38 signaling pathways in zy- mosan-induced down-regulation of CysLT1 expression. A, The iDCs were incubated for 30 min in the absence or presence of SB203580 (SB), U0126 (U), PD98059 (PD), or SP600125 (SP) before addition of zymosan. CysLT1 expression was determined by flow cytometry 48 h later. Data represent geo- metric mean fluorescence intensity (MFI) SEM for n 5 experiments. , p 0.05 and , p 0.02 compared with iDC). B, The iDCs were incubated for 30 min in the absence or presence of SB203580 (SB) or U0126 (U) before addition of zymosan. IL-10 was measured in the culture supernatants by ELISA after 24 h. Data represent mean SEM for n 5 experiments. , p 0.05; , p 0.02; , p 0.01 compared with zymosan.

Article Snippet: MAPK inhibitors U0126 and SP600125 were from Calbiochem; SB203580 was from Sigma-Aldrich; and PD98059 was from InvivoGen.

Techniques: Protein-Protein interactions, Expressing, Incubation, Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Redox biology

Article Title: Notoginsenoside R1-loaded mesoporous silica nanoparticles targeting the site of injury through inflammatory cells improves heart repair after myocardial infarction.

doi: 10.1016/j.redox.2022.102384

Figure Lengend Snippet: Fig. 11. NGR1 protects H9C2 cells from H2O2-induced cell injury through the activation of PI3K/AKT and MAPK/ERK signaling pathways. Changes in intracellular p- AKT and p-ERK1/2 protein levels after treatment of H9C2 cells with 100 μM NGR1 and 350 μM H2O2 (A–C). *P < 0.05 and **P < 0.01 versus the control group; #P < 0.05 and ##P < 0.01 versus the H2O2 group. Changes in intracellular p-AKT and p-ERK levels at various time points in NGR1-treated H9C2 cells (D–G). **P < 0.01 versus the control group; ##P < 0.01 versus the 15 min group; &&P < 0.01 versus the 30 min group. The protective effect of NGR1 on H9C2 cells after the addition of signaling pathway inhibitors (H, I). **P < 0.01 versus the control group; #P < 0.05 and ##P < 0.01 versus the H2O2 group; &P < 0.05 and &&P < 0.01 versus the NGR1 + H2O2 group. TUNEL assay results for each group after the addition of LY294002 and U0126 (J, K). Scale bar: 100 μm **P < 0.01 versus the H2O2 group; #P < 0.05, ##P < 0.01 versus the NGR1 + H2O2 group. Effect of Annexin V-PI flow-through assay LY294002 and U0126 on the protective effect of NGR1 (L, M). **P < 0.01 versus the H2O2 group; #P < 0.05 versus the NGR1 + H2O2 group.

Article Snippet: To explore the mechanisms, H9C2 cells were treated with 25 μM LY294002 (AKT inhibitor, Selleck, USA), 10 μM U0126 (ERK1/2 inhibitor, Selleck, USA) or 5 μM Verteporfin (VP; YAP inhibitor, Selleck, USA) at 30 min before NGR1 treatment.

Techniques: Activation Assay, Protein-Protein interactions, Control, TUNEL Assay

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