Journal: Hypertension

Article Title: Mitogen-Activated Protein Kinases Mediate Upregulation of Hypothalamic Angiotensin II Type 1 Receptors in Heart Failure Rats

doi: 10.1161/hypertensionaha.108.113639

Figure Lengend Snippet: Figure 5. Immunohistochemical analysis of AT1-R immunoreactivity in the PVN of VEH-treated Sham rats and HF rats treated for 4 weeks with ICV VEH, losartan, PD98059, SP600125, or SB203580. A, Representative sections of PVN from animals undergoing each treatment protocol. Third ventricle is to the right. B, Grouped data showing numbers of AT1-R–positive neurons counted in PVN-dp, PVN-mp, PVN-vlp, and PVN-pm. Values are expressed as meansSEM (n6 to 7 for each group). *P0.05 compared with ShamVEH; †P0.05 HFtreatment compared with HFVEH. ‡P0.05, PVN-mp compared with other PVN regions, ShamVEH.

Article Snippet: The selective p44/42 MAPK inhibitor PD98059, the JNK inhibitor SP600125 and the p38 MAPK inhibitor SB203580 were obtained from Tocris (Ellisville, MO).

Techniques: Immunohistochemical staining

Journal: Hypertension

Article Title: Mitogen-Activated Protein Kinases Mediate Upregulation of Hypothalamic Angiotensin II Type 1 Receptors in Heart Failure Rats

doi: 10.1161/hypertensionaha.108.113639

Figure Lengend Snippet: Figure 6. Immunohistochemical analysis of AT1-R immunoreactivity in the SFO of VEH-treated Sham rats and HF rats treated for 4 weeks with ICV VEH, losartan, PD98059, SP600125, or SB203580. A, Representative sections of SFO from animals undergoing each treatment protocol. Third ventricle is at bottom of each image. B, Grouped data showing numbers of AT1-R positive neurons counted in central SFO. Values are expressed as meansSEM (n6 to 7 for each group). *P0.05 compared with ShamVEH; †P0.05 HFtreatment compared with HFVEH.

Article Snippet: The selective p44/42 MAPK inhibitor PD98059, the JNK inhibitor SP600125 and the p38 MAPK inhibitor SB203580 were obtained from Tocris (Ellisville, MO).

Techniques: Immunohistochemical staining

Journal: International journal of molecular sciences

Article Title: MAPK Pathway under Chronic Copper Excess in Green Macroalgae (Chlorophyta): Involvement in the Regulation of Detoxification Mechanisms.

doi: 10.3390/ijms20184546

Figure Lengend Snippet: Figure 7. Principal components ordination (PCO) analysis diagrams in relation with time (a): T1: 6 h, T2: 24 h, T3: 48 h, and T4: 6 d; and treatments (b): t1: Control only with seawater, t2: Solely copper exposure as 10 µM of CuSO4 (Cu), t3: copper + 5 µM MAPK ERK inhibitor PD98059 (Cu + ERKi), t4: Copper + 5 µM MAPK JNK inhibitor SP600125 (Cu + JNKi), t5: Copper + MAPK p38 inhibitor SB203580 (Cu + p38i), t6: Cu + ERKi + JNKi, t7: Cu + ERKi + p38i, t8: Cu + p38i + JNKi, and t9: Cu + ERKi + JNKi + p38i. Vectors overlay (Sperman rank correlation) indicate the relationship between the PCO axes and the parameters H2O2, TBARS (thiobarbituric acid reactive substance), GSH (reduced glutathione), GSSG (oxidised glutathione), ASC (reduced ascorbate), DHA (dehydroascorbate), and relative gene expression catalase (CAT), superoxide dismutase (SOD), dehydroascorbate reductase (DHAR), thioredoxin (TRX), ascorbate peroxidase (APX), and glutathione synthase (GS).

Article Snippet: Nine different treatments, with three replicates each, were conducted as: T1) control with just seawater; T2) only copper exposure as 10 μM of CuSO4 (Sigma-Aldrich, St. Louis, MO, USA) (Cu); T3) copper + 5 μM MAPK ERK inhibitor PD98059 (Tocris Bioscience, St. Louis, MO, USA) (Cu + ERKi); T4) copper + 5 μM MAPK JNK inhibitor SP600125 (Tocris Bioscience, St. Louis, MO, USA) (Cu + JNKi); T5) copper + MAPK p38 inhibitor SB203580 (Tocris Bioscience) (Cu + p38i); T6) Cu + ERKi + JNKi; T7) Cu + ERKi + p38i; T8) Cu + p38i + JNKi; and T9) Cu + ERKi + JNKi + p38i.

Techniques: Control, Gene Expression

Journal: International journal of molecular sciences

Article Title: MAPK Pathway under Chronic Copper Excess in Green Macroalgae (Chlorophyta): Involvement in the Regulation of Detoxification Mechanisms.

doi: 10.3390/ijms20184546

Figure Lengend Snippet: Figure 8. Principal Components Ordination (PCO) analysis diagrams considering the data covered in this article and in the parallel/complementary Celis-Plá et al. [1]. PCOs are related with time (a): T1: 6 h, T2: 24 h, T3: 48 h, and T4: 6 d; and treatments (b): t1: control only with seawater, t2: solely copper exposure as 10 µM CuSO4 (Cu), t3: copper + 5 µM MAPK ERK inhibitor PD98059 (Cu + ERKi), t4: copper + 5 µM MAPK JNK inhibitor SP600125 (Cu + JNKi), T5: copper + MAPK p38 inhibitor SB203580 (Cu + p38i), t6: Cu + ERKi + JNKi, t7: Cu + ERKi + p38i, t8: Cu + p38i + JNKi, and t9: Cu + ERKi + JNKi + p38i. Vectors overlay (Sperman rank correlation) indicate the relationship between the PCO axes and the parameters H2O2, TBARS (thiobarbituric acid reactive substance), GSH (reduced glutathione), GSSG (oxidised glutathione), ASC (reduced ascorbate), DHA (dehydroascorbate), and relative gene expression catalase (CAT), superoxide dismutase (SOD), dehydroascorbate reductase (DHAR), thioredoxin (TRX), ascorbate peroxidase (APX), and glutathione synthase (GS). Furthermore, the physiological variables intracellular copper accumulation (copper photoinhibition (Fv/Fm) productivity (ETRmax), efficiency (αETR) and saturation of irradiance (EkETR), accompanied by higher non-photochemical quenching (NPQmax) [1].

Article Snippet: Nine different treatments, with three replicates each, were conducted as: T1) control with just seawater; T2) only copper exposure as 10 μM of CuSO4 (Sigma-Aldrich, St. Louis, MO, USA) (Cu); T3) copper + 5 μM MAPK ERK inhibitor PD98059 (Tocris Bioscience, St. Louis, MO, USA) (Cu + ERKi); T4) copper + 5 μM MAPK JNK inhibitor SP600125 (Tocris Bioscience, St. Louis, MO, USA) (Cu + JNKi); T5) copper + MAPK p38 inhibitor SB203580 (Tocris Bioscience) (Cu + p38i); T6) Cu + ERKi + JNKi; T7) Cu + ERKi + p38i; T8) Cu + p38i + JNKi; and T9) Cu + ERKi + JNKi + p38i.

Techniques: Control, Gene Expression

Journal: American journal of physiology. Gastrointestinal and liver physiology

Article Title: Functional modulation of enterocytes by gram-positive and gram-negative microorganisms.

doi: 10.1152/ajpgi.00341.2003

Figure Lengend Snippet: Fig. 9. Effects of pretreatment of IEC with MAPK inhibitors on IL-8 secretion and T84 monolayer resistance induced by the PM, EcN, and S. dublin. A: HT-29 and T84 cells were pretreated with 10 M of a p38 MAPK inhibitor (SB-203580) or 50 m of a p42/44 MAPK inhibitor (PD-98059) and subsequently infected with 104 CFU/ml of E. coli or S. dublin. IL-8 secretion in culture supernatants was determined by ELISA. Values represent means SD of n 5 independent assays; *P 0.05; **P 0.01. B: T84 cells were pretreated as indicated above with the MAPK inhibitors. Subsequently, TER was measured at the time points indicated. Results from a representative experiment of 4 independent assays are shown (*P 0.05; **P 0.01). C: medium conditioned by the PM for 12 h was applied to T84 cells for 15 min, and activation of ERK1/2 and p38 was analyzed using phosphospecific antibodies. Reprobing of the blots with antibodies for total ERK or p38 confirmed equal protein loading [lane 1: unstimulated control; lane 2: PM-conditioned medium diluted 1:100; lane 3: PM-conditioned medium diluted 1:10; lane 4: PM-conditioned medium undiluted; lane 5: positive control (10% FCS)]. D: HT-29 cells were pretreated with MAPK inhibitors as indicated above and subsequently stimulated with organisms of the PM for 6 h. Subsequently, mucin gene expression was quantified by real-time PCR. Values represent means SD of n 5 independent assays; *P 0.05.

Article Snippet: Inhibitors of p42/44 mitogen-activated protein kinases (MAPKs; PD-98059) and an inhibitor of p38 MAPK [SB203580 (9)] were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Activation Assay, Control, Positive Control, Gene Expression, Real-time Polymerase Chain Reaction

Journal: Preventive Nutrition and Food Science

Article Title: Kaempferol Inhibits Angiogenesis by Suppressing HIF-1α and VEGFR2 Activation via ERK/p38 MAPK and PI3K/Akt/mTOR Signaling Pathways in Endothelial Cells

doi: 10.3746/pnf.2017.22.4.320

Figure Lengend Snippet: Effect of kaempferol treatment on (A) MAPK and (B) PI3K/Akt/mTOR signaling pathways. Endothelial cells were treated with the indicated concentrations of kaempferol for 24 h. Protein samples (40 μg) were subjected to 10% SDS-PAGE, and phosphorylated ERK, p38, PI3K, Akt, mTOR, p70S6K, VEGFR2, and HIF-1α were detected by western blotting. The cells were treated with kaempferol in combination with MAPK inhibitors (ERK inhibitor, PD98059; p38 inhibitor, SB203580), and the activation of ERK and p38 was detected by western blotting; β-actin was used as an internal control. Values represent the mean±SD of three independent experiments. * P <0.05 and ** P <0.01 compared with control.

Article Snippet: The phospho-specific antibodies (anti-p38, anti-PI3K, anti-AKT, anti-mTOR, anti-p70S6K, anti-VEGFR2, and anti-HIF-1α) and the MAPK inhibitors (SB203580 and PD 98059) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Protein-Protein interactions, SDS Page, Western Blot, Activation Assay, Control