Journal: Frontiers in Cellular Neuroscience

Article Title: Porphyromonas gingivalis -induced periodontitis could contribute to cognitive impairment in Sprague–Dawley rats via the P38 MAPK signaling pathway

doi: 10.3389/fncel.2023.1141339

Figure Lengend Snippet: Effects of periodontitis on the protein level of APP and the activation of the P38 MAPK signaling pathway. (A) All of the above proteins were upregulated by P. gingivalis (except protein P38). High expression of these proteins was effectively inhibited by SB203580. (B–D) The quantification of related protein expression in each group. Experimental values are expressed as the mean ± SEM ( n = 3 per group). ** p < 0.01 compared with the control group, # p < 0.05 compared with the P. gingivalis group.

Article Snippet: We purchased SB203580 (HY-10256A, NJ, USA) from MedChemExpress.

Techniques: Activation Assay, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Differential Inflammatory-Response Kinetics of Human Keratinocytes upon Cytosolic RNA- and DNA-Fragment Induction

doi: 10.3390/ijms19030774

Figure Lengend Snippet: Activation of Nuclear Factor κB (NF-κB), Mitogen Activated Protein Kinase (MAPK) and Signal Transducers of Activator of Transcription (STAT) signal transduction pathways in HPV-KER cells upon poly(I:C) or poly(dA:dT) transfection assessed by NF-κB-luciferase reporter assay ( A ) and western blot analysis ( B – D ). ( A ) NF-κB luciferase reporter assay exhibited faster activation of NF-κB transcription factor upon poly(I:C) treatment than poly(dA:dT) treatment. Raw luminescence intensity values were normalized to the intrinsic control renilla activity, and compared to the 0 h untreated samples. Data are presented as mean of three independent experiments ± standard error; statistical significance was assessed by two-way repeated measurement ANOVA * p < 0.05, grey: poly(I:C) treatment compared to untreated 0 h samples, black: poly(dA:dT) treatment compared to untreated 0 h samples; ( B ) Increase in phosphorylated NF-κB inhibitor α (IκBα) after poly(I:C) or poly(dA:dT) treatment, peaking later after poly(dA:dT) treatment than after poly(I:C) treatment, arrow indicate the lane for phosphorylated IκBα; ( C ) Phosphorylation of ERK1/2 increases after poly(I:C) or poly(dA:dT) treatment, peaking later after poly(dA:dT) treatment than after poly(I:C) treatment, arrows indicate from top to bottom the lanes for phosphorylated ERK1 and ERK2. Phosphorylation of p38 and JNK was not observed upon poly(I:C) or poly(dA:dT) treatment; ( D ) Phosphorylation of both STAT-1 and STAT-3 occurs faster in poly(I:C) treated samples than in poly(dA:dT) treated samples. Western blot results are representative for at least three independent experiments. Actin was used as loading control.

Article Snippet: For inhibition studies, cells were incubated 1 h prior to poly(dA:dT)/poly(I:C) transfection with inhibitors for NF-κB (Bay 11-7085, 10 μM; MedChem Express, Monmouth Junction, NJ, USA), STAT-1 (Fludarabine, 10 μM; Sigma Aldrich), STAT-3 (Stattic, 5 μM; Sigma Aldrich), MEK1 (PD98059, 20 μM; Sigma Aldrich), JNK (SP600125, 10 μM; Tocris Bioscience, Bristol, UK) and p38 (SB203580, 10 μM; Tocris Bioscience).

Techniques: Activation Assay, Transduction, Transfection, Luciferase, Reporter Assay, Western Blot, Control, Activity Assay, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Differential Inflammatory-Response Kinetics of Human Keratinocytes upon Cytosolic RNA- and DNA-Fragment Induction

doi: 10.3390/ijms19030774

Figure Lengend Snippet: Inhibition of different signaling routes has divergent effects on the expression of the IL-6 (white bars) and TNF-α (grey bars) cytokines in keratinocytes. The effect of inhibition by NF-κB ( A ), p38 ( B ), c-Jun N-terminal kinase (JNK) ( C ), mitogen-activated protein kinase kinase 1 and2 (MEK1/2) ( D ), STAT-1 ( E ) and STAT-3 ( F ) on poly(I:C) (6 h after transfection) and poly(dA:dT) (12 h after transfection) induction of IL-6 (white bars) and TNF-α (grey bars) expression in HPV-KER cells. Fold change of mRNA expression values were determined by the ΔΔ C t method, normalized to 18S rRNA expression. As all inhibitors were dissolved in dimethyl sulfoxide (DMSO), the relative mRNA expression levels were compared to the expression levels in samples treated with DMSO + poly(I:C) or DMSO + poly(dA:dT), respectively. Poly(I:C) and poly(dA:dT) induction was in every case significant compared to the untreated control samples; no significant difference was observed between the cytokine-expression level of the samples treated with poly(I:C), poly(dA:dT), DMSO + poly(I:C) or DMSO + poly(dA:dT). Data are represented as the means of three independent experiments ± standard error; * p < 0.05; ** p < 0.01; *** p < 0.001 determined by Student’s t -test.

Article Snippet: For inhibition studies, cells were incubated 1 h prior to poly(dA:dT)/poly(I:C) transfection with inhibitors for NF-κB (Bay 11-7085, 10 μM; MedChem Express, Monmouth Junction, NJ, USA), STAT-1 (Fludarabine, 10 μM; Sigma Aldrich), STAT-3 (Stattic, 5 μM; Sigma Aldrich), MEK1 (PD98059, 20 μM; Sigma Aldrich), JNK (SP600125, 10 μM; Tocris Bioscience, Bristol, UK) and p38 (SB203580, 10 μM; Tocris Bioscience).

Techniques: Inhibition, Expressing, Transfection, Control

Journal: Journal of Endocrinology

Article Title: Angiotensin II enhances the increase in monocyte chemoattractant protein-1 production induced by tumor necrosis factor-α from 3T3-L1 preadipocytes

doi: 10.1677/joe-08-0363

Figure Lengend Snippet: Figure 3 Effect of ERK1/2 inhibitor (PD98059), p38MAPK inhibitor (SB203580), or NF-kB inhibitor (TPCK) on Ang II and/or TNF-a- induced MCP-1 mRNA expression in 3T3-L1 preadipocytes. 3T3-L1 preadipocytes pretreated with PD98059 (20 mM), SB203580 (20 mM), or TPCK (20 mM) for 1 h were stimulated with Ang II (1 mM) and/or TNF-a (1 ng/ml) for 2 h for measurement of MCP-1 mRNA levels using real-time RT-PCR. Data were obtained from three independent experiments and expressed as a fold increase over control. *P!0.05, **P!0.005.

Article Snippet: After that, Ang II and/or TNF-a were added to the medium and incubated for 3 h. A specific inhibitor of mitogen-activated protein kinase (MAPK) ERK kinase, PD98059 (Cell Signaling Technology) and a specific inhibitor of p38MAPK, SB203580 (Cell Signaling Technology), and NF-kB inhibitor N-tosyll-phenylalanine-chloromethyl ketone (TPCK, Sigma– Aldrich Corp.) were added to the medium 1 h before stimulation with Ang II and/or TNF-a.

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Toxicology Reports

Article Title: Emissions from commercial-grade charbroiling meat operations induce oxidative stress and inflammatory responses in human bronchial epithelial cells

doi: 10.1016/j.toxrep.2014.09.015

Figure Lengend Snippet: The role of p38 in mediating the pro-inflammatory effect of OE. (A) Increased p38 phosphorylation in OE-treated BEAS-2B cells. Cells were exposed to OE for 2 and 4 h. Total p38 was used as internal control. (B) Effect of p38 inhibitor on IL-6 production by OE-treated cells. (C). Effect of p38 inhibitor on OE-induced IL-8 production. BEAS-2B cells were pre-incubated with p38 inhibitor SB203580 for 2 h prior to being exposed to OE for 16 h. OE50: OE at 50 μg/ml. Values represent means ± SEM, n = 3, * p < 0.05 compared to the respective control; ** p < 0.05 compared to OE alone.

Article Snippet: R,S-Sulforaphane (SFN) and SB203580 were purchased from LKT Laboratories (St. Paul, MN) and EMD Millipore (Bedford, MA), respectively.

Techniques: Phospho-proteomics, Control, Incubation

Journal: Toxicology Reports

Article Title: Emissions from commercial-grade charbroiling meat operations induce oxidative stress and inflammatory responses in human bronchial epithelial cells

doi: 10.1016/j.toxrep.2014.09.015

Figure Lengend Snippet: Up-regulation of COX-2 and PGE2 by OE. (A) Increased COX-2 protein in BEAS-2B cells treated with OE for 16 h. (B) Effects of NAC (1 mM), SFN (5 μM) and SB203580 (5 μM) on PGE2 release from BEAS-2B cells after16-h exposure to 50 μg/ml of OE. Values represent means ± SEM, n = 3. * p < 0.05 compared to the control; ** p < 0.05 compared to OE alone.

Article Snippet: R,S-Sulforaphane (SFN) and SB203580 were purchased from LKT Laboratories (St. Paul, MN) and EMD Millipore (Bedford, MA), respectively.

Techniques: Control

Journal: The Journal of biological chemistry

Article Title: Cell volume-dependent regulation of L-selectin shedding in neutrophils. A role for p38 mitogen-activated protein kinase.

doi: 10.1074/jbc.274.31.22072

Figure Lengend Snippet: FIG. 7. The role of p38 mitogen-activated protein kinase in the shedding of L-selectin induced by hypertonicity and other stim- uli. A, neutrophils were treated with either genistein (40 mM) or PP1 (10 mM) for 20 min and then challenged with hypertonicity (100 mM extra NaCl) for 10 min or 2 h. Cells were lysed, subjected to SDS-PAGE, and immunoblotted using polyclonal anti-phospho-p38. The same blot was then stripped and reprobed with an anti-p38 antibody. B, following the same treatment as in A and exposure to hypertonicity for 2 h, p38 was immunoprecipitated with a specific anti-p38 antibody. In vitro kinase assay was then performed using ATF-2 as a substrate, and the results were visualized with autoradiography. C, neutrophils were treated in the presence or absence of the p38 inhibitor SB203580 (20 mM) for 20 min. The medium was then supplemented with an extra 100 mM NaCl for 2 h (HT), LPS (1 mg/ml) for 1 h, FMLP (100 nM), or PMA (50 nM) both for 20 min. L-selectin surface expression was measured using a mAb and flow cytometry. The data represent the means 6 S.E., n 5 6 separate studies. *, p , 0.001 versus control (CON); **, p , 0.05 versus control; ***, p , 0.01 versus control; Genist., genistein; HT, hypertonic medium.

Article Snippet: FMLP, PMA, urea, sucrose, lipopolysaccharide (Escherichia coli O111: B4), bovine serum albumin, diisopropylfluorophosphate, and propidium iodide were from Sigma, and Triton X-100 was from Caledon Lab. BCECF was obtained from Molecular Probes Inc.; genistein, erbstatin analog, SB203580, DEVD-fmk, and staurosporin were from Calbiochem; the proteinase inhibitor mixture was from PharMingen; and ATF-2 was from Santa Cruz Biotechnology.

Techniques: SDS Page, Immunoprecipitation, In Vitro, Kinase Assay, Autoradiography, Expressing, Flow Cytometry, Control

Journal: Cardiovascular research

Article Title: Gadd45γ regulates cardiomyocyte death and post-myocardial infarction left ventricular remodelling.

doi: 10.1093/cvr/cvv219

Figure Lengend Snippet: Figure 7 Gadd45g promotes cardiomyocyte death in a p38 MAPK-, RIP1-, and caspase-8-dependent manner. (A) Representative images showing cardiomyocyte nuclei positive for TUNEL in neonatal cardiomyocytes with or without Gadd45g overexpression in the presence of scrambled siRNA, RIP1 siRNA, caspase-8 siRNA, or SB203580 (10 mM). (B) Quantification showing the number of TUNEL-positive myocyte nuclei as a per cent of total nuclei (DAPI staining) after Gadd45g overexpression (n ¼ 9). (C) Representative immunoblots showing the expression pattern of p-p38, T-p38, p-HSP27, cleaved caspase-8, cleaved RIP1, cleaved caspase-3, Gadd45g, GAPDH, and Ponceau S staining in neonatal cardiomyocytes subjected to Gadd45g overexpression with or without 10 mM of SB203580 (n ¼ 6–9). Values are mean + SEM. (D) Representative immunoblots showing the ex- pression pattern of caspase-8, cleaved caspase-3, Gadd45g, GAPDH, and Ponceau S staining in neonatal cardiomyocytes subjected to Gadd45g over- expression with or without casapse-8 siRNA (n ¼ 6–9). (E) Representative immunoblots showing the expression pattern of RIP1, cleaved caspase-3, Gadd45g, GAPDH, and Ponceau S staining in neonatal cardiomyocytes subjected to Gadd45g overexpression with or without RIP1 siRNA (n ¼ 6–9).

Article Snippet: All chemicals were purchased from Sigma Aldrich Chimie (France) except for SB203580 hydrochloride (Tocris, UK, catalogue no. 1402).

Techniques: TUNEL Assay, Over Expression, Staining, Western Blot, Expressing