mapk erk inhibitor pd98059  (Cell Signaling Technology Inc)

Journal: Antioxidants

Article Title: NADPH Oxidase 5 (NOX5) Upregulates MMP-10 Production and Cell Migration in Human Endothelial Cells

doi: 10.3390/antiox13101199

Figure Lengend Snippet: p38 MAPK, MEK/ERK, and JNK pathways participate at different levels in NOX5-dependent regulation of MMP-10: ( A ) MMP-10 mRNA levels of tHMock and tHNOX5 cell lines after 24 h of incubation with 5 μM 219138-24-6 (p38 MAPK inhibitor, ip38), PD98059 (MEK/ERK inhibitor, iERK), or JNK-IN-8 (JNK inhibitor, iJNK) ( n = 6). ( B ) MMP-10 protein in the conditioned medium of tHMock and tHNOX5 cell lines after 24 h of incubation with 5 μM 219138-24-6 (ip38), PD98059 (iERK), or JNK-IN-8 (iJNK) ( n = 6). tHMock: stable cell line transfected with pcDNA3.2-Mock. tHNOX5: stable cell line transfected with pcDNA3.2-NOX5. n.s.: not significant differences, * p < 0.05, *** p < 0.001, **** p < 0.0001. Data are presented as median and IQR.

Article Snippet: The p38 MAPK inhibitor (ab145872, Abcam ® , Waltham, MA, USA), JNK MAPK inhibitor (JNK-IN-8, SML1246, Sigma Aldrich), and ERK MAPK inhibitor (PD98059, 9900S, Cell Signaling Technology, Danvers, MA, USA) were used at final concentrations of 5 μM in the cell medium.

Techniques: Incubation, Stable Transfection, Transfection

Journal: Antioxidants

Article Title: NADPH Oxidase 5 (NOX5) Upregulates MMP-10 Production and Cell Migration in Human Endothelial Cells

doi: 10.3390/antiox13101199

Figure Lengend Snippet: NOX5 enhances MMP-10 promoter activity via the JNK pathway and a functional AP-1 site: ( A ) MMP-10 promoter activity of tHMock and tHNOX5 cell lines at baseline and stimulated with 0.25 μM Ang II ( n = 6). ( B ) MMP-10 promoter activity of tHMock and tHNOX5 cell lines after 24 h of incubation with 5 μM 219138-24-6 (p38 MAPK inhibitor, ip38), PD98059 (MEK/ERK inhibitor, iERK), or JNK-IN-8 (JNK inhibitor, iJNK) ( n = 6). ( C ) Schematic representation of MMP-10 promoter constructions used and quantification of MMP-10 promoter activity of tHMock and tHNOX5 cell lines transfected with MMP-10 promoter constructions ( n = 6). CREB: cAMP response element binding protein (CREB) putative union site. AP-1: activator protein-1 (AP-1) putative union site. Crosses indicate site-directed mutations in the putative union sites. tHMock: stable cell line transfected with pcDNA3.2-Mock. tHNOX5: stable cell line transfected with pcDNA3.2-NOX5. ns: not significant differences, * p < 0.05 vs. tHMock Ctrl, *** p < 0.001, **** p < 0.0001, # p < 0.05 vs. tHNOX5 Ctrl cells. Data are presented as median and IQR or as mean ± SEM.

Article Snippet: The p38 MAPK inhibitor (ab145872, Abcam ® , Waltham, MA, USA), JNK MAPK inhibitor (JNK-IN-8, SML1246, Sigma Aldrich), and ERK MAPK inhibitor (PD98059, 9900S, Cell Signaling Technology, Danvers, MA, USA) were used at final concentrations of 5 μM in the cell medium.

Techniques: Activity Assay, Functional Assay, Incubation, Transfection, Binding Assay, Stable Transfection

Journal: Journal of inflammation (London, England)

Article Title: Combination of IL-17A/F and TNF-α uniquely alters the bronchial epithelial cell proteome to enhance proteins that augment neutrophil migration.

doi: 10.1186/s12950-022-00323-w

Figure Lengend Snippet: Fig. 5 Assessment of pharmacological inhibitors on IL-17A/F and TNF-α mediated production of selected proteins. HBEC-3KT cells were pre-treated with pharmacological inhibitors (A) LY294002 (PI3Ki), (B) GO6976 (PKCi) and (C) PD98059 (MEKi), for 1 h prior to stimulation with IL-17A/F (50 ng/ ml), TNF-α (20 ng/ml) or the combination of IL-17A/F and TNF-α. Tissue culture supernatants were collected after 24 h and examined for protein production by ELISA for LCN-2 and Elafin. Protein abundance shown is after subtraction of background abundance in paired unstimulated cells in each independent replicate. Each data point represents an independent experimental replicate (n = 4) and the line represents the average. Two-way ANOVA with Dunnett’s test for multiple comparisons was used to determine statistical significance (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001)

Article Snippet: Pharmacological inhibitors, phosphoinositide 3-kinase (PI3K) inhibitor LY294002, protein kinase-C (PKC) inhibitor GO6976 and MAPK/ERK kinase (MEK) inhibitor PD98059 were obtained from SelleckChem (Burlington, ON, CA).

Techniques: Enzyme-linked Immunosorbent Assay, Quantitative Proteomics

Journal: Molecular Vision

Article Title: NGF promotes cell cycle progression by regulating D-type cyclins via PI3K/Akt and MAPK/Erk activation in human corneal epithelial cells

doi:

Figure Lengend Snippet: Nerve growth factor promotes cell cycle progression via Akt and Erk activation in human corneal epithelial cells. A : Human corneal epithelial cells (HCECs) were incubated in keratinocyte serum-free medium (K-SFM) without growth factors for 24 h before treatment with nerve growth factor (NGF) at 25 ng/ml for 1 h. A total of 50 µg cell lysates and total RNA were analyzed for expression of the indicated genes by western blot analysis and reverse transcriptase–polymerase chain reaction (RT–PCR), respectively. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. B : HCECs were incubated in K-SFM without growth factors for 24 h and then pretreated with LY294002 or PD98059 at 10 µM for 1 h before treatment with NGF at 25 ng/ml for another hour. A total of 50 μg cell lysates were analyzed for expression of the indicated genes by immunoblotting analysis. GAPDH was used as a loading control. Experiments were performed in triplicate. C : The schematic representation depicts how the Akt and Erk pathways collaborate to control NGF induction of cyclin D expression.

Article Snippet: Drugs were used as follows: human recombinant NGF-β from Sigma-Aldrich (St. Louis, MO), diluted in PBS containing 0.1% BSA; the PI3K/Akt inhibitor LY294002; and the MAPK/ Erk inhibitor PD98059 from Sigma-Aldrich, diluted in dimethyl sulfoxide.

Techniques: Activation Assay, Incubation, Expressing, Western Blot, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction