mapk kinase inhibitors pd98059  (Millipore)

Journal: Biomedicines

Article Title: SGLT2 Inhibitor Canagliflozin Alleviates High Glucose-Induced Inflammatory Toxicity in BV-2 Microglia.

doi: 10.3390/biomedicines12010036

Figure Lengend Snippet: Figure 5. Cana suppresses the induction of iNOS and the secretion of inflammatory cytokines IL-1β and TNF-α in HG-treated BV-2 cells but augments the expression of COX-2. (a,b) NG-cultured cells, with or without the four SGLT2 inhibitors (10 and 20 µM) given 30 min in advance, were stimulated with HG (50 mM) for 24 h. Cultures treated with HG served as the control group. The iNOS and COX-2 levels were analyzed using Western blotting. The relatively densitometric analysis results were shown using histograms, and values are expressed as mean ± SEM (n = 4). # p < 0.05 vs. HG group’s iNOS; & p < 0.05 vs. HG group’s COX-2. (c) NG-cultured cells, with or without specific signaling pathway inhibitors (20 µM PD98059 (PD), 10 µM SP600125 (SP), 15 µM SB203580 (SB), 10 µM LY294002 (LY), and 3 µM CAPE) given 30 min in advance were exposed to 50 mM HG for 24 h. After Western blotting detection, the relatively densitometric analysis results of iNOS and COX-2 levels were shown using histograms, and values are expressed as mean ± SEM (n = 4). # p < 0.05 vs. HG group’s iNOS; & p < 0.05 vs. HG group’s COX-2. (d,e) The same treatments as (a,b) were administered, and the cultures were incubated only for 12 h. The secretion levels of IL-1β and TNF-α were assayed using ELISA. Results were shown using histograms and values are expressed as mean ± SEM (n = 4, in triplicate). # p < 0.05 vs. HG group.

Article Snippet: Chemical agents, including mitogen-activated protein kinase (MAPK) inhibitor PD98059, c-Jun N-terminal kinase (JNK) inhibitor SP600125, p38 MAPK inhibitor SB203580, and phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, were purchased from Selleckchem (Houston, TX, USA).

Techniques: Expressing, Cell Culture, Control, Western Blot, Incubation, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Cerebroprotective Effect of 17β-Estradiol Replacement Therapy in Ovariectomy-Induced Post-Menopausal Rats Subjected to Ischemic Stroke: Role of MAPK/ERK1/2 Pathway and PI3K-Independent Akt Activation

doi: 10.3390/ijms241814303

Figure Lengend Snippet: The MEK inhibitor PD98059, but not the PI3K inhibitor LY294002, attenuates estradiol protection. Infarct sizes in brains from placebo- and E2-treated ovariectomized female rats subjected to tMCAO. ( A ) Representative images of the TTC-stained third coronal brain slice from placebo- (−) and E2-treated (+) animals, treated with vehicle, PD98059 or LY294002. ( B ) Summary data showing total infarct volumes, expressed as a percentage of the corresponding non-ischemic hemisphere. Two-way ANOVA for estrogen treatment (E2 vs. placebo, p < 0.0001, F 1,31 = 32.5), and inhibitors treatment (PD98059 and LY294002 vs. vehicle, p < 0.01, F 2,31 = 7.9). Post hoc Sidak’s multiple comparisons tests: significantly different from placebo group (* p < 0.05 and ** p < 0.01) or from vehicle-treated group ( # p < 0.05). Data are mean ± SEM of individual data points. E2, 17β-estradiol. tMCAO, transient middle cerebral artery occlusion.

Article Snippet: Anesthetized animals were treated with intracerebroventricular (icv) administration of the selective MAPK kinase (MEK) inhibitor PD98059 (3 µg, Calbiochem, La Jolla, CA, USA), the potent and specific PI3K inhibitor LY294002 (30 µg, Promega; Madison, WI) or vehicle (10 µL of 50% DMSO).

Techniques: Staining, Slice Preparation

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