Journal: Obesity (Silver Spring, Md.)

Article Title: IGF-I attenuates FFA-induced activation of JNK1 phosphorylation and TNFα expression in human subcutaneous preadipocytes.

doi: 10.1002/oby.20329

Figure Lengend Snippet: FIGURE 3 FFA and MALP-2, but not Pam3CSK4 increase phosphorylation of JNK. Human SC preadipocytes were serum-starved overnight and treated with Pam3CSK4 1 or 10 lg/ml, MALP-2 2 or 5 lg/ml, or FFAs 0.1 mM for 20 min fol- lowed by WB analysis of cell lysates using phospho and total JNK, ERK and p38 antibodies. Representative blot is shown; this was repeated twice with similar results.

Article Snippet: Mitogen activated protein kinase (MAPK) inhibitors for ERK, p38, and JNK were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Phospho-proteomics

Journal: Obesity (Silver Spring, Md.)

Article Title: IGF-I attenuates FFA-induced activation of JNK1 phosphorylation and TNFα expression in human subcutaneous preadipocytes.

doi: 10.1002/oby.20329

Figure Lengend Snippet: FIGURE 5 IGF-I decreases FFA and MALP-2-stimulated JNK phosphorylation. Human SC preadipocytes were serum-starved overnight and then pretreated with 10 nM IGF-I for 1 h followed by control, MALP-2 5lg/ml, or FFAs 0.1 mM for 20 min. Total cell lysates were analyzed by WB using phospho and total JNK and p38 antibodies. A representative blot is shown and data are presented as mean þ SE of densi- tometry analyses of fold over basal (phospho-JNK and phospho-p38 corrected for total JNK and total p38) of three experiments; *P < 0.05.

Article Snippet: Mitogen activated protein kinase (MAPK) inhibitors for ERK, p38, and JNK were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Phospho-proteomics, Control

Journal: The Journal of Biological Chemistry

Article Title: Transcription of the Transforming Growth Factor ? Activating Integrin ?8 Subunit Is Regulated by SP3, AP-1, and the p38 Pathway *

doi: 10.1074/jbc.M110.113977

Figure Lengend Snippet: p38 regulates ITGB8 expression and αvβ8-mediated TGF-β activation. A, flow cytometry for integrin β8 on adult lung fibroblasts treated with MAPK inhibitors, PD98059 (ERK), SB202190 (p38), and SP600125 (JNK) (± S.E.). MFI, mean fluorescence intensity. B, quantitative RT-PCR results for ITGB8 expression in adult lung fibroblasts treated with SB202190, normalized to GAPDH and β-actin and relative to control (± S.E.). C, immunoblot for phosphorylated HSP 27 and dual-phosphorylated ATF-2 from nuclear extracts from adult lung fibroblasts treated ± SB202190. Immunoblot for the nuclear localized proteins, lamins A and C, was used as a loading control. D, TGF-β activation assays of adult lung fibroblasts treated with anti-β8 blocking antibodies or SB202190 (± S.E.). E, quantitative RT-PCR results for MAPK14 (p38α) and ITGB8 in adult lung fibroblasts transfected with plasmids expressing a p38α dominant-negative isoform (p38αDN) or the empty vector control, pcDNA (± S.E.). The measured transcript is labeled above each respective graph. F, TGF-β activation assays of adult lung fibroblasts transfected with plasmids expressing a p38α dominant-negative isoform (p38αDN) or the empty vector control, pcDNA. Percentage (%) of αvβ8-mediated TGF-β activation shown (± S.E.). * = p ≤ 0.05; ** = p ≤ 0.01; *** = p ≤ 0.001.

Article Snippet: The chemical inhibitors of the p38 (SB202190) and extracellular-signaling response kinase (ERK) (PD98059) MAP kinase pathways were obtained from Calbiochem (EMD4 Biosciences, Gibbstown, NJ), whereas the inhibitor of the Jun N-terminal kinase (JNK) pathway (SP600125) was obtained from A.G. Scientific, Inc. (San Diego, CA).

Techniques: Expressing, Activation Assay, Flow Cytometry, Fluorescence, Quantitative RT-PCR, Western Blot, Blocking Assay, Transfection, Dominant Negative Mutation, Plasmid Preparation, Labeling

Journal: PLoS ONE

Article Title: IL-17 Enhances Chemotaxis of Primary Human B Cells during Asthma

doi: 10.1371/journal.pone.0114604

Figure Lengend Snippet: (A) B cells isolated from asthmatic patients were pre-treated with different kinase inhibitors or DMSO carrier for 1 hr before being incubated with Th-17 cytokines in a migration assay. Data is presented as % migration of B cells relative to negative control (medium). (n = 6) (B) Western blots showing p38 MAPK phosphorylation in B cells following Th-17 cytokine stimulation. Asthmatic B cells (2×10 6 ) stimulated with Th-17 cytokines were lysed using 1x RIPA buffer and cell lysates were resolved using western analysis. Blots were probed using anti-phospho-p38 and anti-p38 MAPK antibodies. (C) Ratio of p-p38 over p38 band intensity were determined using densitometer and data were presented as fold increase in p38 phosphorylation ratio following Th-17 cytokine stimulation compared to non-stimulated condition (medium). Data is expressed as means ± SD (n = 5). *P<0.05 is considered significant.

Article Snippet: Specifically, we used the p38 MAPK inhibitor SB203580 (0.1 mM; Axon Medchem, Groningen, The Netherlands), the extracellular signal-regulated kinase (ERK) 1/2 MAPK inhibitor PD184352 (2 mM; United States Biological, Inc, Swampscott, Mass), the NF-kB inhibitor PS1145 (10 mM; Professor Sir Philip Cohen), and the phosphoinositide 3-kinase (PI3K) inhibitor PI103 (5 mM; Cayman Chemical, Ann Arbor, Mich).

Techniques: Isolation, Incubation, Migration, Negative Control, Western Blot, Phospho-proteomics

Journal: Journal of Nanobiotechnology

Article Title: Intracellular signaling pathway in dendritic cells and antigen transport pathway in vivo mediated by an OVA@DDAB/PLGA nano-vaccine

doi: 10.1186/s12951-021-01116-8

Figure Lengend Snippet: Cytokine mRNA expression and activation of relevant signal pathway profiles elicited by different vaccine adjuvants for BMDCs. a , b , c The cytokine (IL-1β, IFN-γ and TNF-α) mRNA expression from BMDCs was analyzed by RT-qPCR. d The changes of p38 MAPK, p-AKT and p-ERK phosphorylation after being stimulated with different formulation DCs for 6 h and 12 h. e Change of the p38 MAPK, p-AKT and p-ERK phosphorylation level by stimulated DCs with different concentration of DDAB-PLGA Nv. f DDAB-PLGA Nv increased the binding of MKK3 to its substrate p38α. g The ability of AP2α to recruit TNF-α and IL-1β promoters in the P38 pathway. h Illustration of how OVA@DDAB/PLGA Nv promoted DCs activation and maturation via the p38 MAPK signaling pathway. i After treatment with inhibitors of p38 (SB203580), the change of OVA@DDAB/PLGA Nv induced cytokine secretion from BMDCs. j After treatment with inhibitors of p38 (SB203580), the change of OVA@DDAB/PLGA Nv induced cytokine mRNA expression from BMDCs. Data are expressed as Mean ± STD (n = 6). (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)

Article Snippet: OVA derived (H-2 Kb, SIINFEKL) specific MHC I pentamers was purchased from ProImmune (Oxford, UK). p38 MAPK inhibitor SB203580 was purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Concentration Assay, Binding Assay