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  • 99
    Name:
    Stat1 Antibody
    Description:
    The Stat1 transcription factor is activated in response to a large number of ligands 1 and is essential for responsiveness to IFN α and IFN γ 2 3 Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization nuclear translocation and DNA binding 4 Stat1 protein exists as a pair of isoforms Stat1α 91 kDa and the splice variant Stat1β 84 kDa In most cells both isoforms are activated by IFN α but only Stat1α is activated by IFN γ The inappropriate activation of Stat1 occurs in many tumors 5 In addition to tyrosine phosphorylation Stat1 is also phosphorylated at Ser727 through a p38 mitogen activated protein kinase MAPK dependent pathway in response to IFN α and other cellular stresses 6 Serine phosphorylation may be required for the maximal induction of Stat1 mediated gene activation
    Catalog Number:
    9172
    Price:
    None
    Applications:
    Western Blot, Immunoprecipitation, Chromatin Immunoprecipitation
    Category:
    Primary Antibodies
    Source:
    Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human Stat1. Antibodies are purified by protein A and peptide affinity chromatography.
    Reactivity:
    Human Mouse Rat Monkey
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc p38
    Dose‐dependent reduction of age‐related cardiac inflammation by phenolic compounds. (a) Representative microphotographs of left ventricular sections of the heart stained with hematoxylin‐eosin showing young, SHAM, DMSO, PC 2.5, PC 5, PC 10, and PC 20 groups. (b) Histograms showing cardiomyocytes width (μm) and semiquantitative scores of inflammation ( n = 6–8 per group). (c) CRP and IL‐6 plasma concentrations in all groups ( n = 6–8 per group). (d) Representative western blot analyses for <t>p38</t> in rat cardiac tissue, normalized to GAPDH (in arbitrary units, a.u.) ( n = 3 for each protein and condition). * p
    The Stat1 transcription factor is activated in response to a large number of ligands 1 and is essential for responsiveness to IFN α and IFN γ 2 3 Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization nuclear translocation and DNA binding 4 Stat1 protein exists as a pair of isoforms Stat1α 91 kDa and the splice variant Stat1β 84 kDa In most cells both isoforms are activated by IFN α but only Stat1α is activated by IFN γ The inappropriate activation of Stat1 occurs in many tumors 5 In addition to tyrosine phosphorylation Stat1 is also phosphorylated at Ser727 through a p38 mitogen activated protein kinase MAPK dependent pathway in response to IFN α and other cellular stresses 6 Serine phosphorylation may be required for the maximal induction of Stat1 mediated gene activation
    https://www.bioz.com/result/p38/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1216 article reviews
    Price from $9.99 to $1999.99
    p38 - by Bioz Stars, 2020-10
    99/100 stars

    Images

    1) Product Images from "Long‐term intake of phenolic compounds attenuates age‐related cardiac remodeling, et al. Long‐term intake of phenolic compounds attenuates age‐related cardiac remodeling"

    Article Title: Long‐term intake of phenolic compounds attenuates age‐related cardiac remodeling, et al. Long‐term intake of phenolic compounds attenuates age‐related cardiac remodeling

    Journal: Aging Cell

    doi: 10.1111/acel.12894

    Dose‐dependent reduction of age‐related cardiac inflammation by phenolic compounds. (a) Representative microphotographs of left ventricular sections of the heart stained with hematoxylin‐eosin showing young, SHAM, DMSO, PC 2.5, PC 5, PC 10, and PC 20 groups. (b) Histograms showing cardiomyocytes width (μm) and semiquantitative scores of inflammation ( n = 6–8 per group). (c) CRP and IL‐6 plasma concentrations in all groups ( n = 6–8 per group). (d) Representative western blot analyses for p38 in rat cardiac tissue, normalized to GAPDH (in arbitrary units, a.u.) ( n = 3 for each protein and condition). * p
    Figure Legend Snippet: Dose‐dependent reduction of age‐related cardiac inflammation by phenolic compounds. (a) Representative microphotographs of left ventricular sections of the heart stained with hematoxylin‐eosin showing young, SHAM, DMSO, PC 2.5, PC 5, PC 10, and PC 20 groups. (b) Histograms showing cardiomyocytes width (μm) and semiquantitative scores of inflammation ( n = 6–8 per group). (c) CRP and IL‐6 plasma concentrations in all groups ( n = 6–8 per group). (d) Representative western blot analyses for p38 in rat cardiac tissue, normalized to GAPDH (in arbitrary units, a.u.) ( n = 3 for each protein and condition). * p

    Techniques Used: Staining, Western Blot

    2) Product Images from "Nitrogen mustard exposure of murine skin induces DNA damage, oxidative stress and activation of MAPK/Akt-AP1 pathway leading to induction of inflammatory and proteolytic mediators"

    Article Title: Nitrogen mustard exposure of murine skin induces DNA damage, oxidative stress and activation of MAPK/Akt-AP1 pathway leading to induction of inflammatory and proteolytic mediators

    Journal: Toxicology letters

    doi: 10.1016/j.toxlet.2015.04.006

    Effect of NM on the phosphorylation of MAPK signaling molecules in SKH-1 hairless mouse skin Mice were exposed topically to either 3.2 mg NM in acetone or acetone alone and dorsal skin was collected at 12, 24, 72 and 120 h time points after exposure. Skin tissue lysates were prepared and subjected to western blot analysis as detailed under ‘Materials and Methods’ Section 2. The membranes were probed for phospho- and total ERK1/2 (A), JNK1/2 (B) and p38 (C). Protein loading was checked by stripping and re-probing the membranes with β-actin antibody. Results obtained were quantified by densitometric analysis of representative immunoblots for phospho and total ERK1/2, JNK1/2 and p38. Results shown are representative of 2–3 animals in the study group. VC, vehicle (acetone) control; NM, nitrogen mustard.
    Figure Legend Snippet: Effect of NM on the phosphorylation of MAPK signaling molecules in SKH-1 hairless mouse skin Mice were exposed topically to either 3.2 mg NM in acetone or acetone alone and dorsal skin was collected at 12, 24, 72 and 120 h time points after exposure. Skin tissue lysates were prepared and subjected to western blot analysis as detailed under ‘Materials and Methods’ Section 2. The membranes were probed for phospho- and total ERK1/2 (A), JNK1/2 (B) and p38 (C). Protein loading was checked by stripping and re-probing the membranes with β-actin antibody. Results obtained were quantified by densitometric analysis of representative immunoblots for phospho and total ERK1/2, JNK1/2 and p38. Results shown are representative of 2–3 animals in the study group. VC, vehicle (acetone) control; NM, nitrogen mustard.

    Techniques Used: Mouse Assay, Western Blot, Stripping Membranes

    3) Product Images from "Differences in the Signaling Pathways of ?1A- and ?1B-Adrenoceptors Are Related to Different Endosomal Targeting"

    Article Title: Differences in the Signaling Pathways of ?1A- and ?1B-Adrenoceptors Are Related to Different Endosomal Targeting

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0064996

    α 1A - and α 1B -AR stimulation shows a subtype-specific pattern of p38-MAPK phosphorylation. HEK293 cells stably transfected with VSV-tagged or untagged α 1A - and α 1B -subtypes were serum-starved for 4 hours and stimulated with PHE (100 µM) for a 15 minute time-course at 37°C. In some experiments, prazosin (10 µM) or 5-methylurapidil (10 µM) were added for 30 min. After stimulation, cellular extracts were prepared as described under the “Experimental procedures”. Equal amounts (15 µg) of each sample were used to visualize the phosphorylated p-38 MAPK expression Western blots from representative experiments are shown. The lower panels show total p38 loaded on each sample. The graph quantifies the p-p38 MAPK signal at different times. Data represent means ± S.E.M. of 3-6 independent experiments. Student’s t t est was applied to determine significant differences at a given time vs control, *p
    Figure Legend Snippet: α 1A - and α 1B -AR stimulation shows a subtype-specific pattern of p38-MAPK phosphorylation. HEK293 cells stably transfected with VSV-tagged or untagged α 1A - and α 1B -subtypes were serum-starved for 4 hours and stimulated with PHE (100 µM) for a 15 minute time-course at 37°C. In some experiments, prazosin (10 µM) or 5-methylurapidil (10 µM) were added for 30 min. After stimulation, cellular extracts were prepared as described under the “Experimental procedures”. Equal amounts (15 µg) of each sample were used to visualize the phosphorylated p-38 MAPK expression Western blots from representative experiments are shown. The lower panels show total p38 loaded on each sample. The graph quantifies the p-p38 MAPK signal at different times. Data represent means ± S.E.M. of 3-6 independent experiments. Student’s t t est was applied to determine significant differences at a given time vs control, *p

    Techniques Used: Stable Transfection, Transfection, Expressing, Western Blot

    α 1A - and α 1B -AR mediated p-38 activation is modulated by methyl-β-cyclodextrin, filipin and concanavalin A. HEK293 cells stably transfected with each α 1 -AR subtype were serum-starved for 4 hours and stimulated with PHE (100 µM) for a 15 minute time-course at 37°C. In some experiments, methyl-β-cyclodextrin 10 mM (mβCD) was added for 30 or 60 min, filipin 1 µg/ml and concanavalin A 250 µg/ml for 30 min. After stimulation, cellular extracts were prepared as described under the “Experimental procedures”. Equal amounts (15 µg) of each sample were used to visualize the p-p38-MAPK expression Western blots from representative experiments are shown. The lower panels show amounts of p38-MAPK loaded on each sample. The graph quantifies the p-p38-MAPK signal at different times after agonist addition. Data represent means ± S.E.M. of 3–4 independent experiments. Student’s t t est was applied to determine significant differences at a given time vs control, ***p
    Figure Legend Snippet: α 1A - and α 1B -AR mediated p-38 activation is modulated by methyl-β-cyclodextrin, filipin and concanavalin A. HEK293 cells stably transfected with each α 1 -AR subtype were serum-starved for 4 hours and stimulated with PHE (100 µM) for a 15 minute time-course at 37°C. In some experiments, methyl-β-cyclodextrin 10 mM (mβCD) was added for 30 or 60 min, filipin 1 µg/ml and concanavalin A 250 µg/ml for 30 min. After stimulation, cellular extracts were prepared as described under the “Experimental procedures”. Equal amounts (15 µg) of each sample were used to visualize the p-p38-MAPK expression Western blots from representative experiments are shown. The lower panels show amounts of p38-MAPK loaded on each sample. The graph quantifies the p-p38-MAPK signal at different times after agonist addition. Data represent means ± S.E.M. of 3–4 independent experiments. Student’s t t est was applied to determine significant differences at a given time vs control, ***p

    Techniques Used: Activation Assay, Stable Transfection, Transfection, Expressing, Western Blot

    4) Product Images from "Cellular Changes during Renal Failure-Induced Inflammatory Aortic Valve Disease"

    Article Title: Cellular Changes during Renal Failure-Induced Inflammatory Aortic Valve Disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0129725

    Macrophages osteoblast markers and intracellular pathways in early phases of calcification. (A) Immunostaining of CD68 (I), osteopontin (II), and osteocalcin (III) were positive in the valve annulus (An) and leaflets (Le) after 2 weeks on the nephropathic diet. (B) Western blot analysis (n = 3 in each group) of osteocalcin, osteopontin, and Runx-2 (I), and of the ratio of phosphorylated ERK to ERK-1(II). There were no changes in the expression Akt, JNK, or p38 pathways (III). Graphic presentation of Western blot analysis (IV).
    Figure Legend Snippet: Macrophages osteoblast markers and intracellular pathways in early phases of calcification. (A) Immunostaining of CD68 (I), osteopontin (II), and osteocalcin (III) were positive in the valve annulus (An) and leaflets (Le) after 2 weeks on the nephropathic diet. (B) Western blot analysis (n = 3 in each group) of osteocalcin, osteopontin, and Runx-2 (I), and of the ratio of phosphorylated ERK to ERK-1(II). There were no changes in the expression Akt, JNK, or p38 pathways (III). Graphic presentation of Western blot analysis (IV).

    Techniques Used: Immunostaining, Western Blot, Expressing

    5) Product Images from "MKP1 mediates chemosensitizer effects of E1a in response to cisplatin in non-small cell lung carcinoma cells"

    Article Title: MKP1 mediates chemosensitizer effects of E1a in response to cisplatin in non-small cell lung carcinoma cells

    Journal: Oncotarget

    doi:

    Upregulation of MKP1 mediates E1a associated sensitivity to cDDP in a p38MAPK dependent fashion ( A ) 50 μg of the TCL from NSCLC expressing E1a 13s or empty vector were used to evaluate the MKP1, P-p38MAPK and p38MAPK. Tubulin was used as loading control. ( B ) H1299 stably transfected with pcDNA E1a 13s (H1299 E1a) or empty vector (E.V.) were infected with lentivirus carrying sh-RNA against MKP1 (H1299 E1a shMKP1) and then mRNA MKP1 levels were evaluated by qRT-PCR. ( C ) 50 μg of TCL from cells used in B) were blotted against MKP1 and E1a. Tubulin was used as loading control. ( D ) 50 μg of TCL from cell used in C) were blotted against p38, ERK1/2 and the respective active forms. Tubulin was used as loading control. ( E ) H1299 E.V, H1299 E1a and H1299 E1a and sh-RNA MKP1 were treated with the indicated doses of cDDP during 48 hours and viability was evaluated by crystal violet method. Bars indicate standard deviation (SD). ( F ) H1299 E.V and H1299 E1a were treated with the indicated doses of cDDP during 48 hours in the presence/absences of PD98059 (10 μM). Viability was evaluated by crystal violet method. Bars indicate standard deviation (SD). ( G ) H1299 E.V and H1299 E1a were treated with the indicated doses of cDDP during 48 hours in the presence/absences of SB203580 (10 μM). Viability was evaluated by crystal violet method. Bars indicate standard deviation (SD).
    Figure Legend Snippet: Upregulation of MKP1 mediates E1a associated sensitivity to cDDP in a p38MAPK dependent fashion ( A ) 50 μg of the TCL from NSCLC expressing E1a 13s or empty vector were used to evaluate the MKP1, P-p38MAPK and p38MAPK. Tubulin was used as loading control. ( B ) H1299 stably transfected with pcDNA E1a 13s (H1299 E1a) or empty vector (E.V.) were infected with lentivirus carrying sh-RNA against MKP1 (H1299 E1a shMKP1) and then mRNA MKP1 levels were evaluated by qRT-PCR. ( C ) 50 μg of TCL from cells used in B) were blotted against MKP1 and E1a. Tubulin was used as loading control. ( D ) 50 μg of TCL from cell used in C) were blotted against p38, ERK1/2 and the respective active forms. Tubulin was used as loading control. ( E ) H1299 E.V, H1299 E1a and H1299 E1a and sh-RNA MKP1 were treated with the indicated doses of cDDP during 48 hours and viability was evaluated by crystal violet method. Bars indicate standard deviation (SD). ( F ) H1299 E.V and H1299 E1a were treated with the indicated doses of cDDP during 48 hours in the presence/absences of PD98059 (10 μM). Viability was evaluated by crystal violet method. Bars indicate standard deviation (SD). ( G ) H1299 E.V and H1299 E1a were treated with the indicated doses of cDDP during 48 hours in the presence/absences of SB203580 (10 μM). Viability was evaluated by crystal violet method. Bars indicate standard deviation (SD).

    Techniques Used: Expressing, Plasmid Preparation, Stable Transfection, Transfection, Infection, Quantitative RT-PCR, Standard Deviation

    6) Product Images from "Disease modifying effect of adiponectin in model of α-synucleinopathies"

    Article Title: Disease modifying effect of adiponectin in model of α-synucleinopathies

    Journal: Annals of Clinical and Translational Neurology

    doi: 10.1002/acn3.77

    APN ameliorates neurodegeneration in a cell model of α -synucleinopathies. B103 neuroblastoma cells expressing human α S or empty vector were treated with rec. APN (5μg/ml) or PBS. Cells were fractionated into TBS-, SDS-, and FA-extractable fractions) 16 (A–C) and analyzed by immunoblotting using anti- α S. Uncropped blots of (a) are presented in Figure S4. In (C), cells were pretransfected with siRNA of AdipoRI or nontarget (control), while in (C) cells were preincubated with a p38 inhibitor SB203580 (1 μ mol/L) or a AMPK inhibitor compound C (1 μ mol/L). In (D–F), phosphorylation of α S was evaluated by immunoblotting (D and F) or immunofluoresence (E) using anti -pα S or anti- α S. In (E), representative image of double immunofluorescence showed that colocalization of pα S with α S was reduced by APN. In (F), cells were pretransfected with siRNA for AdipoRI or nontarget. In (G), the suppressive effect of APN on release of α S in the conditioned medium (CM) was evaluated, while exosomes were semipurified from the CM and analyzed for α S and two exosome markers: alix and flotillin-1. The intensities of the immunoreactivities of α S were quantified (A: all fractions, B and C: FA fraction, mean ± SEM, n = 3–5, n.s; not significant, * P
    Figure Legend Snippet: APN ameliorates neurodegeneration in a cell model of α -synucleinopathies. B103 neuroblastoma cells expressing human α S or empty vector were treated with rec. APN (5μg/ml) or PBS. Cells were fractionated into TBS-, SDS-, and FA-extractable fractions) 16 (A–C) and analyzed by immunoblotting using anti- α S. Uncropped blots of (a) are presented in Figure S4. In (C), cells were pretransfected with siRNA of AdipoRI or nontarget (control), while in (C) cells were preincubated with a p38 inhibitor SB203580 (1 μ mol/L) or a AMPK inhibitor compound C (1 μ mol/L). In (D–F), phosphorylation of α S was evaluated by immunoblotting (D and F) or immunofluoresence (E) using anti -pα S or anti- α S. In (E), representative image of double immunofluorescence showed that colocalization of pα S with α S was reduced by APN. In (F), cells were pretransfected with siRNA for AdipoRI or nontarget. In (G), the suppressive effect of APN on release of α S in the conditioned medium (CM) was evaluated, while exosomes were semipurified from the CM and analyzed for α S and two exosome markers: alix and flotillin-1. The intensities of the immunoreactivities of α S were quantified (A: all fractions, B and C: FA fraction, mean ± SEM, n = 3–5, n.s; not significant, * P

    Techniques Used: Expressing, Plasmid Preparation, Immunofluorescence

    7) Product Images from "5‐ALA/SFC enhances HO‐1 expression through the MAPK/Nrf2 antioxidant pathway and attenuates murine tubular epithelial cell apoptosis"

    Article Title: 5‐ALA/SFC enhances HO‐1 expression through the MAPK/Nrf2 antioxidant pathway and attenuates murine tubular epithelial cell apoptosis

    Journal: FEBS Open Bio

    doi: 10.1002/2211-5463.12729

    5‐ALA/SFC increased HO‐1 and Nrf2 expression via activation of MAPK signaling pathway in mProx24 cells. (A) After pretreatment with HO‐1 siRNA, representative bands of different groups showed HO‐1, Nrf2, p38, p‐p38, Erk‐1/2, p‐Erk‐1/2, Bcl‐2, and cleaved‐caspase‐3 proteins in mProx24 cells treated with 5‐ALA/SFC. (B) Quantitative densitometry was performed. Data are the mean ± SD. Statistical significance was measured by Student's t ‐test. Correlations were determined by Spearman's ranking ( n = 3; * P
    Figure Legend Snippet: 5‐ALA/SFC increased HO‐1 and Nrf2 expression via activation of MAPK signaling pathway in mProx24 cells. (A) After pretreatment with HO‐1 siRNA, representative bands of different groups showed HO‐1, Nrf2, p38, p‐p38, Erk‐1/2, p‐Erk‐1/2, Bcl‐2, and cleaved‐caspase‐3 proteins in mProx24 cells treated with 5‐ALA/SFC. (B) Quantitative densitometry was performed. Data are the mean ± SD. Statistical significance was measured by Student's t ‐test. Correlations were determined by Spearman's ranking ( n = 3; * P

    Techniques Used: Expressing, Activation Assay

    8) Product Images from "Type 2 diabetes‐induced hyposalivation of the submandibular gland through PINK1/Parkin‐mediated mitophagy, et al. Type 2 diabetes‐induced hyposalivation of the submandibular gland through PINK1/Parkin‐mediated mitophagy"

    Article Title: Type 2 diabetes‐induced hyposalivation of the submandibular gland through PINK1/Parkin‐mediated mitophagy, et al. Type 2 diabetes‐induced hyposalivation of the submandibular gland through PINK1/Parkin‐mediated mitophagy

    Journal: Journal of Cellular Physiology

    doi: 10.1002/jcp.28962

    ERK1/2 is required for HG‐induced mitophagy in SMG‐C6 cells. The levels of phosphorylated and total ERK1/2 (a and b), JNK (c and d) and p38 (e,f) were detected by western blot analysis. β‐Actin was used as a loading control. n = 6, * p
    Figure Legend Snippet: ERK1/2 is required for HG‐induced mitophagy in SMG‐C6 cells. The levels of phosphorylated and total ERK1/2 (a and b), JNK (c and d) and p38 (e,f) were detected by western blot analysis. β‐Actin was used as a loading control. n = 6, * p

    Techniques Used: Western Blot

    9) Product Images from "Characterization and allergic role of IL-33-induced neutrophil polarization"

    Article Title: Characterization and allergic role of IL-33-induced neutrophil polarization

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/cmi.2017.163

    The specific phenotype of IL-33-treated neutrophils is mediated by JNK and NF-κB signaling pathways. ( a ) Neutrophils freshly isolated from bone marrow were treated with IL-33 for 15, 30 and 120 min, respectively. The activities of JNK, p38, STAT1, Erk and p65 in cytoplasm were detected by western blotting. ( b ) The c-jun and p65 protein levels in nucleus of neutrophils after IL-33 treatment for 1 and 2 h, respectively, were determined by western blotting. ( c and d ) Neutrophils were pretreated with the indicated concentrations of JNK inhibitor (SP600125) and NF-κB inhibitor for 30 min and then stimulated with IL-33 for 24 h. The levels of IL-9, CCL2, CCL7, CCL12, IL-1R2 and IL-13Ra1 expression were detected by real-time PCR. ( e and f ) The concentrations of IL-9, IL-4, IL-5, IL-13, CCL2 and CCL7 in the supernatant of neutrophils pretreated with either SP600125 or NF-κB inhibitor and then cultured with IL-33 for 24 h were determined by ELISA assay kits. Assays were performed more than three times. Data are shown as mean±s.d. ( n =3). * P
    Figure Legend Snippet: The specific phenotype of IL-33-treated neutrophils is mediated by JNK and NF-κB signaling pathways. ( a ) Neutrophils freshly isolated from bone marrow were treated with IL-33 for 15, 30 and 120 min, respectively. The activities of JNK, p38, STAT1, Erk and p65 in cytoplasm were detected by western blotting. ( b ) The c-jun and p65 protein levels in nucleus of neutrophils after IL-33 treatment for 1 and 2 h, respectively, were determined by western blotting. ( c and d ) Neutrophils were pretreated with the indicated concentrations of JNK inhibitor (SP600125) and NF-κB inhibitor for 30 min and then stimulated with IL-33 for 24 h. The levels of IL-9, CCL2, CCL7, CCL12, IL-1R2 and IL-13Ra1 expression were detected by real-time PCR. ( e and f ) The concentrations of IL-9, IL-4, IL-5, IL-13, CCL2 and CCL7 in the supernatant of neutrophils pretreated with either SP600125 or NF-κB inhibitor and then cultured with IL-33 for 24 h were determined by ELISA assay kits. Assays were performed more than three times. Data are shown as mean±s.d. ( n =3). * P

    Techniques Used: Isolation, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay

    10) Product Images from "β-Phenethyl Isothiocyanate Induces Cell Death in Human Osteosarcoma through Altering Iron Metabolism, Disturbing the Redox Balance, and Activating the MAPK Signaling Pathway"

    Article Title: β-Phenethyl Isothiocyanate Induces Cell Death in Human Osteosarcoma through Altering Iron Metabolism, Disturbing the Redox Balance, and Activating the MAPK Signaling Pathway

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2020/5021983

    PEITC suppressed OS growth in vivo . MNNG/HOS cells were injected subcutaneously into the right flank of the male BALB/c nude mice. One week after the OS xenograft mouse model was established, the mice were randomly divided into two groups and, respectively, administrated with 10% sesame seed oil and 30 mg/kg PEITC once daily for 24 consecutive days. (a) Body weight change of the two groups. (b) Volume change of OS tissues of the two groups. Data were calculated by the formula: volume = length × width 2 × 0.5. (c) Weight of OS tissues from the two groups. (d) H E staining analysis of tumor tissues (200× and 400×). (e) The protein expression levels of TfR1, DMT1, FTH1, and FPN in tumor tissues. (f) The protein expression levels of LC3B, C-caspase3, GPx4, and Cdc2 in tumor tissues. (g) Phosphorylation levels of ERK, p38, and JNK in tumor tissues. All data were presented as the means ± SD ( n = 7). ∗ P
    Figure Legend Snippet: PEITC suppressed OS growth in vivo . MNNG/HOS cells were injected subcutaneously into the right flank of the male BALB/c nude mice. One week after the OS xenograft mouse model was established, the mice were randomly divided into two groups and, respectively, administrated with 10% sesame seed oil and 30 mg/kg PEITC once daily for 24 consecutive days. (a) Body weight change of the two groups. (b) Volume change of OS tissues of the two groups. Data were calculated by the formula: volume = length × width 2 × 0.5. (c) Weight of OS tissues from the two groups. (d) H E staining analysis of tumor tissues (200× and 400×). (e) The protein expression levels of TfR1, DMT1, FTH1, and FPN in tumor tissues. (f) The protein expression levels of LC3B, C-caspase3, GPx4, and Cdc2 in tumor tissues. (g) Phosphorylation levels of ERK, p38, and JNK in tumor tissues. All data were presented as the means ± SD ( n = 7). ∗ P

    Techniques Used: In Vivo, Injection, Mouse Assay, Staining, Expressing

    PEITC induced cell death via ROS generation in human OS cells. (a) EdU staining assay of MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC for 24 h. (b) Quantitative analysis of EdU staining in (a). (c) Protein expression levels of TfR1, DMT1, FTH1, and FPN in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC for 24 h. (d) Protein expression levels of GPx4, LC3B, C-PARP, C-caspase3, Bcl2, and Cdc2 in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC 24 h. (e) Phosphorylation levels of ERK, p38, and JNK in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC for 24 h.
    Figure Legend Snippet: PEITC induced cell death via ROS generation in human OS cells. (a) EdU staining assay of MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC for 24 h. (b) Quantitative analysis of EdU staining in (a). (c) Protein expression levels of TfR1, DMT1, FTH1, and FPN in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC for 24 h. (d) Protein expression levels of GPx4, LC3B, C-PARP, C-caspase3, Bcl2, and Cdc2 in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC 24 h. (e) Phosphorylation levels of ERK, p38, and JNK in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with 30 μ M PEITC in the presence or absence of NAC for 24 h.

    Techniques Used: Staining, Expressing

    PEITC activated the MAPK signaling pathway in human OS cells. (a–d) Phosphorylation levels of ERK, p38, and JNK in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with the indicated concentrations of PEITC for 20 h or 30 μ M PEITC for 4 h, 12 h, 24 h, and 48 h.
    Figure Legend Snippet: PEITC activated the MAPK signaling pathway in human OS cells. (a–d) Phosphorylation levels of ERK, p38, and JNK in MNNG/HOS, U-2 OS, MG-63, and 143B cells treated with the indicated concentrations of PEITC for 20 h or 30 μ M PEITC for 4 h, 12 h, 24 h, and 48 h.

    Techniques Used:

    11) Product Images from "Activation of Cytosolic Phospholipase A2α in Resident Peritoneal Macrophages by Listeria monocytogenes Involves Listeriolysin O and TLR2"

    Article Title: Activation of Cytosolic Phospholipase A2α in Resident Peritoneal Macrophages by Listeria monocytogenes Involves Listeriolysin O and TLR2

    Journal:

    doi: 10.1074/jbc.M709956200

    Activation of ERKs and p38 by WTLM and Δ hly LM contributes to cPLA 2 α -mediated AA release
    Figure Legend Snippet: Activation of ERKs and p38 by WTLM and Δ hly LM contributes to cPLA 2 α -mediated AA release

    Techniques Used: Activation Assay

    TLR2 regulates p38 and ERK activation in macrophages infected with WTLM and Δ hly LM
    Figure Legend Snippet: TLR2 regulates p38 and ERK activation in macrophages infected with WTLM and Δ hly LM

    Techniques Used: Activation Assay, Infection

    12) Product Images from "Plumbagin Inhibits Proliferative and Inflammatory Responses of T Cells Independent of ROS Generation But by Modulating Intracellular Thiols"

    Article Title: Plumbagin Inhibits Proliferative and Inflammatory Responses of T Cells Independent of ROS Generation But by Modulating Intracellular Thiols

    Journal: Journal of cellular biochemistry

    doi: 10.1002/jcb.22620

    Inhibition of proliferation/survival associated signaling molecules by plumbagin and their modulation by GSH in activated T cells. Lymphocytes were incubated with plumbagin (1 µM, 4 h) in presence or absence of GSH and were stimulated with con A (5 µg/ml) for 1 h (A,C) or 24 h (B). Whole cell lysates were prepared, fractionated on 10% SDS–PAGE, and electrotransferred to nitrocellulose membrane. Western blot analysis was performed using different antibodies specific for (A) P-ERK, ERK, P-IKK-α/β, IKK-α, IKK-β, IκB-α, (B) BCL-2, BCL-xL, Cyclin A, and (C) P-P38, P-38, P-JNK, JNK, P-AKT, AKT. β-Actin was used as loading control. Two such independent experiments were carried out.
    Figure Legend Snippet: Inhibition of proliferation/survival associated signaling molecules by plumbagin and their modulation by GSH in activated T cells. Lymphocytes were incubated with plumbagin (1 µM, 4 h) in presence or absence of GSH and were stimulated with con A (5 µg/ml) for 1 h (A,C) or 24 h (B). Whole cell lysates were prepared, fractionated on 10% SDS–PAGE, and electrotransferred to nitrocellulose membrane. Western blot analysis was performed using different antibodies specific for (A) P-ERK, ERK, P-IKK-α/β, IKK-α, IKK-β, IκB-α, (B) BCL-2, BCL-xL, Cyclin A, and (C) P-P38, P-38, P-JNK, JNK, P-AKT, AKT. β-Actin was used as loading control. Two such independent experiments were carried out.

    Techniques Used: Inhibition, Incubation, SDS Page, Western Blot

    13) Product Images from "Rotavirus Activates JNK and p38 Signaling Pathways in Intestinal Cells, Leading to AP-1-Driven Transcriptional Responses and Enhanced Virus Replication ▿"

    Article Title: Rotavirus Activates JNK and p38 Signaling Pathways in Intestinal Cells, Leading to AP-1-Driven Transcriptional Responses and Enhanced Virus Replication ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00390-06

    Effects of JNK and p38 inhibition on RRV structural antigen expression. Caco-2 cells were mock infected or infected with RRV at an MOI of 0.1 (A) or 1.0 (B) in the presence of DMSO, the JNK inhibitor SP600125 (JNKi; 20 μM) or the p38 inhibitor SB203580 (p38i; 50 μM). At 16 h PI, cells were stained with rabbit polyclonal antibodies to RRV and analyzed by flow cytometry as described in Materials and Methods.
    Figure Legend Snippet: Effects of JNK and p38 inhibition on RRV structural antigen expression. Caco-2 cells were mock infected or infected with RRV at an MOI of 0.1 (A) or 1.0 (B) in the presence of DMSO, the JNK inhibitor SP600125 (JNKi; 20 μM) or the p38 inhibitor SB203580 (p38i; 50 μM). At 16 h PI, cells were stained with rabbit polyclonal antibodies to RRV and analyzed by flow cytometry as described in Materials and Methods.

    Techniques Used: Inhibition, Expressing, Infection, Staining, Flow Cytometry, Cytometry

    Effects of JNK and p38 inhibition on RRV replication. (A) HT-29, Caco-2, and MA104 cells were mock infected (M) or infected with RRV at an MOI of 10 in the presence of DMSO alone (−) or the JNK inhibitor SP600125 (JNKi) at the indicated concentrations. Cell lysates were prepared at 8 h PI (HT-29 and MA104) or 16 h PI (Caco-2) and analyzed by Western blotting using antibodies against phosphorylated c-Jun (p-c-Jun). Blots were stripped and reprobed with antibodies to actin as a loading control. HT-29 (B), Caco-2 (C), and MA104 (D) cells infected with RRV at an MOI of 0.1 were incubated in the presence of SP600125 (JNKi), SB203580 (p38i), PD98059 (MEKi), or DMSO alone (D). The infectious titer of virus harvested at 17 h PI was determined as described in Materials and Methods. The data are shown as the mean and standard deviation of triplicate samples from an experiment representative of the two to three independent experiments performed.
    Figure Legend Snippet: Effects of JNK and p38 inhibition on RRV replication. (A) HT-29, Caco-2, and MA104 cells were mock infected (M) or infected with RRV at an MOI of 10 in the presence of DMSO alone (−) or the JNK inhibitor SP600125 (JNKi) at the indicated concentrations. Cell lysates were prepared at 8 h PI (HT-29 and MA104) or 16 h PI (Caco-2) and analyzed by Western blotting using antibodies against phosphorylated c-Jun (p-c-Jun). Blots were stripped and reprobed with antibodies to actin as a loading control. HT-29 (B), Caco-2 (C), and MA104 (D) cells infected with RRV at an MOI of 0.1 were incubated in the presence of SP600125 (JNKi), SB203580 (p38i), PD98059 (MEKi), or DMSO alone (D). The infectious titer of virus harvested at 17 h PI was determined as described in Materials and Methods. The data are shown as the mean and standard deviation of triplicate samples from an experiment representative of the two to three independent experiments performed.

    Techniques Used: Inhibition, Infection, Western Blot, Incubation, Standard Deviation

    Kinetics of p38 activation following RRV infection. Lysates prepared from HT-29 (A), Caco-2 (B), and MA104 (C) cells that had been mock infected or infected with RRV or I-RRV at an MOI of 10 for the indicated times were analyzed by Western blotting using antibodies against active phosphorylated p38 (p-p38). Blots were subsequently stripped and reprobed with antibodies recognizing all forms of p38 (p38).
    Figure Legend Snippet: Kinetics of p38 activation following RRV infection. Lysates prepared from HT-29 (A), Caco-2 (B), and MA104 (C) cells that had been mock infected or infected with RRV or I-RRV at an MOI of 10 for the indicated times were analyzed by Western blotting using antibodies against active phosphorylated p38 (p-p38). Blots were subsequently stripped and reprobed with antibodies recognizing all forms of p38 (p38).

    Techniques Used: Activation Assay, Infection, Western Blot

    Effect of RRV MOI on JNK and p38 activation. Lysates prepared from HT-29 (A), Caco-2 (B), and MA104 (C) cells infected with RRV at the indicated MOI were analyzed at 8 h PI (HT-29 and MA104) and 16 h PI (Caco-2) by Western blotting using antibodies against active phosphorylated JNK (p-JNK) or active phosphorylated p38 (p-p38). Gels were stripped and reprobed with antibodies against all forms of JNK (JNK) or p38 (p38).
    Figure Legend Snippet: Effect of RRV MOI on JNK and p38 activation. Lysates prepared from HT-29 (A), Caco-2 (B), and MA104 (C) cells infected with RRV at the indicated MOI were analyzed at 8 h PI (HT-29 and MA104) and 16 h PI (Caco-2) by Western blotting using antibodies against active phosphorylated JNK (p-JNK) or active phosphorylated p38 (p-p38). Gels were stripped and reprobed with antibodies against all forms of JNK (JNK) or p38 (p38).

    Techniques Used: Activation Assay, Infection, Western Blot

    JNK and p38 activation by multiple rotavirus strains. HT-29 and Caco-2 cells were infected with rotavirus strains RRV, SA11, CRW-8, Wa, and UK at an MOI of 10 or mock infected (M). (A) Cell lysates were prepared at 8 h PI (HT-29) or 16 h PI (Caco-2) and analyzed by Western blotting using antibodies against active phosphorylated JNK (p-JNK) and p38 (p-p38). Blots were subsequently stripped and reprobed with antibody recognizing all forms of JNK (JNK) and p38 (p38). (B) Lysates from cells infected with Wa and UK or mock-infected cells (M) were prepared at 16 h PI (HT-29) or 24 h PI (Caco-2) and analyzed by Western blotting as described for panel A.
    Figure Legend Snippet: JNK and p38 activation by multiple rotavirus strains. HT-29 and Caco-2 cells were infected with rotavirus strains RRV, SA11, CRW-8, Wa, and UK at an MOI of 10 or mock infected (M). (A) Cell lysates were prepared at 8 h PI (HT-29) or 16 h PI (Caco-2) and analyzed by Western blotting using antibodies against active phosphorylated JNK (p-JNK) and p38 (p-p38). Blots were subsequently stripped and reprobed with antibody recognizing all forms of JNK (JNK) and p38 (p38). (B) Lysates from cells infected with Wa and UK or mock-infected cells (M) were prepared at 16 h PI (HT-29) or 24 h PI (Caco-2) and analyzed by Western blotting as described for panel A.

    Techniques Used: Activation Assay, Infection, Western Blot

    Role of JNK and p38 in RRV-induced gene expression. HT-29 cells were mock infected or infected with RRV or I-RRV at an MOI of 10 for 8 h in the presence of DMSO alone (−), SP600125 (JNKi), or SB203580 (p38i). Cellular RNA was extracted and analyzed by real-time PCR for levels of IL-8 mRNA induced by RRV (A) and I-RRV (B) and c- jun mRNA induced by RRV (C). The data are shown as the mean and standard deviation of triplicate samples from an experiment representative of the two independent experiments performed.
    Figure Legend Snippet: Role of JNK and p38 in RRV-induced gene expression. HT-29 cells were mock infected or infected with RRV or I-RRV at an MOI of 10 for 8 h in the presence of DMSO alone (−), SP600125 (JNKi), or SB203580 (p38i). Cellular RNA was extracted and analyzed by real-time PCR for levels of IL-8 mRNA induced by RRV (A) and I-RRV (B) and c- jun mRNA induced by RRV (C). The data are shown as the mean and standard deviation of triplicate samples from an experiment representative of the two independent experiments performed.

    Techniques Used: Expressing, Infection, Real-time Polymerase Chain Reaction, Standard Deviation

    14) Product Images from "A GFP-based assay for rapid screening of compounds affecting ARE-dependent mRNA turnover"

    Article Title: A GFP-based assay for rapid screening of compounds affecting ARE-dependent mRNA turnover

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gnh086

    Western blots of 15-GFP–IL3 lysates treated with ionomycin, okadaic acid and MG132 for 30 min. Activation of the p38 MAPK pathway was determined with an antibody directed against the phosphorylated, active form of p38. JNK pathway activation
    Figure Legend Snippet: Western blots of 15-GFP–IL3 lysates treated with ionomycin, okadaic acid and MG132 for 30 min. Activation of the p38 MAPK pathway was determined with an antibody directed against the phosphorylated, active form of p38. JNK pathway activation

    Techniques Used: Western Blot, Activation Assay

    15) Product Images from "Type I IFN induces protein ISGylation to enhance cytokine expression and augments colonic inflammation"

    Article Title: Type I IFN induces protein ISGylation to enhance cytokine expression and augments colonic inflammation

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1505690112

    ISGylation of ubiquitylated proteins in MEFs delays their processing by proteasomes, and protein ISGylation facilitates p38 activation in DSS-treated colon. ( A ) Total protein ISGylation and ubiquitylation in whole lysates of WT and Ube1L-KO MEFs were analyzed by immunoblotting. MEFs were treated with IFN for 24 h and with MG132 for an additional 8 h. ( B ) Cellular ubiquitylated proteins were isolated from an equal amount of protein lysates of WT and Ube1L-KO MEFs by the ubiquitin-binding protein HR23A. Protein on beads was immunoblotted for ubiquitylation and ISGylation by ubiquitin- and ISG15-specific antibodies, respectively. One representative of three experiments is shown. H. C., heavy chain. L. C., light chain. ( C ) Isolated ubiquitylated proteins from WT and Ube1L-KO MEFs were incubated with 26S proteasomes for the indicated times. The processing of ubiquitylated proteins was visualized by ubiquitin antibody. The relative amounts of Flag-HR23A in the isolated proteins were detected with the anti-Flag blot. Data are shown from one representative experiment of two independent experiments. ( D ) The intensity of signals from ubiquitylated proteins was measured from the Western blot shown in C using Li-Cor analysis software. The total ubiquitylated protein at time 0 was set as 1, and the others were normalized by the total protein at time 0 in each group. Data are presented as mean + SD of two independent experiments. P , student’s t -test. ( E ) Cell lysates of colons from DSS-treated WT/Ube1L-KO mice were analyzed by SDS/PAGE, followed by immunoblotting with antibodies as indicated. Three mice in each cohort were analyzed. ( F ) The intensity of signals from phosphorylation of p38 and polyubiquitylated proteins was measured from the Western blots shown in E using Li-Cor analysis software. The signal intensity of one Ube1L-KO sample was set as 1, and the others were normalized accordingly. Data are presented as mean + SEM ( n = 3). P , student’s t -test.
    Figure Legend Snippet: ISGylation of ubiquitylated proteins in MEFs delays their processing by proteasomes, and protein ISGylation facilitates p38 activation in DSS-treated colon. ( A ) Total protein ISGylation and ubiquitylation in whole lysates of WT and Ube1L-KO MEFs were analyzed by immunoblotting. MEFs were treated with IFN for 24 h and with MG132 for an additional 8 h. ( B ) Cellular ubiquitylated proteins were isolated from an equal amount of protein lysates of WT and Ube1L-KO MEFs by the ubiquitin-binding protein HR23A. Protein on beads was immunoblotted for ubiquitylation and ISGylation by ubiquitin- and ISG15-specific antibodies, respectively. One representative of three experiments is shown. H. C., heavy chain. L. C., light chain. ( C ) Isolated ubiquitylated proteins from WT and Ube1L-KO MEFs were incubated with 26S proteasomes for the indicated times. The processing of ubiquitylated proteins was visualized by ubiquitin antibody. The relative amounts of Flag-HR23A in the isolated proteins were detected with the anti-Flag blot. Data are shown from one representative experiment of two independent experiments. ( D ) The intensity of signals from ubiquitylated proteins was measured from the Western blot shown in C using Li-Cor analysis software. The total ubiquitylated protein at time 0 was set as 1, and the others were normalized by the total protein at time 0 in each group. Data are presented as mean + SD of two independent experiments. P , student’s t -test. ( E ) Cell lysates of colons from DSS-treated WT/Ube1L-KO mice were analyzed by SDS/PAGE, followed by immunoblotting with antibodies as indicated. Three mice in each cohort were analyzed. ( F ) The intensity of signals from phosphorylation of p38 and polyubiquitylated proteins was measured from the Western blots shown in E using Li-Cor analysis software. The signal intensity of one Ube1L-KO sample was set as 1, and the others were normalized accordingly. Data are presented as mean + SEM ( n = 3). P , student’s t -test.

    Techniques Used: Activation Assay, Isolation, Binding Assay, Incubation, Western Blot, Software, Mouse Assay, SDS Page

    Protein ISGylation promotes IFN-mediated cytokine production through the ROS-p38 axis. ( A ) ELISA of TNF-α in culture supernatants of WT and Ube1L-KO BMDMs incubated with LPS (100 ng/mL) for the indicated time after IFN priming for 24 h. Representative data are presented as mean + SD from two independent experiments. P , student’s t -test. ( B ) Immunoblot analysis of IkBα, total and phosphorylated (p-) p38, and JNK in lysates of WT and Ube1L-KO BMDMs stimulated for 0–120 min (times are shown above lanes) with LPS (100 ng/mL) after IFN priming for 24 h. β-Actin served as a loading control. Data are shown for one representative experiment of three independent experiments. ( C ) Phosphorylation of p38 in LPS-treated WT/Ube1L-KO BMDMs. After IFN priming for 24 h, BMDMs were incubated with LPS (100 ng/mL) for the indicated time. Phosphorylation of p38 was detected by Western blotting. The signal intensity of phosphorylated p38 in Ube1L-KO cells at 30 min was set as 1, and others were normalized accordingly and are shown as mean + SD ( n = 3). P , student’s t -test. ( D ) WT and Ube1L-KO BMDMs were treated with 500 U/mL type I IFN for indicated time period. Cellular ROS was measured by DCFDA staining. Representative data from two independent experiments are shown. Data are presented as mean + SD ( n = 4). P , student’s t -test. N.S., not significant. ( E ) Immunoblot analysis of cellular ISGylation in WT/Ube1L-KO MEFs after IFN treatment for the indicated time. β-Actin served as a loading control. ( F ) WT and Ube1L-KO MEFs were treated with 1,000 U/mL type I IFN for the indicated time period. Cellular ROS was determined, and representative data from triplet assays of two independent experiments are shown as mean + SD ( n = 3). P , student’s t -test.
    Figure Legend Snippet: Protein ISGylation promotes IFN-mediated cytokine production through the ROS-p38 axis. ( A ) ELISA of TNF-α in culture supernatants of WT and Ube1L-KO BMDMs incubated with LPS (100 ng/mL) for the indicated time after IFN priming for 24 h. Representative data are presented as mean + SD from two independent experiments. P , student’s t -test. ( B ) Immunoblot analysis of IkBα, total and phosphorylated (p-) p38, and JNK in lysates of WT and Ube1L-KO BMDMs stimulated for 0–120 min (times are shown above lanes) with LPS (100 ng/mL) after IFN priming for 24 h. β-Actin served as a loading control. Data are shown for one representative experiment of three independent experiments. ( C ) Phosphorylation of p38 in LPS-treated WT/Ube1L-KO BMDMs. After IFN priming for 24 h, BMDMs were incubated with LPS (100 ng/mL) for the indicated time. Phosphorylation of p38 was detected by Western blotting. The signal intensity of phosphorylated p38 in Ube1L-KO cells at 30 min was set as 1, and others were normalized accordingly and are shown as mean + SD ( n = 3). P , student’s t -test. ( D ) WT and Ube1L-KO BMDMs were treated with 500 U/mL type I IFN for indicated time period. Cellular ROS was measured by DCFDA staining. Representative data from two independent experiments are shown. Data are presented as mean + SD ( n = 4). P , student’s t -test. N.S., not significant. ( E ) Immunoblot analysis of cellular ISGylation in WT/Ube1L-KO MEFs after IFN treatment for the indicated time. β-Actin served as a loading control. ( F ) WT and Ube1L-KO MEFs were treated with 1,000 U/mL type I IFN for the indicated time period. Cellular ROS was determined, and representative data from triplet assays of two independent experiments are shown as mean + SD ( n = 3). P , student’s t -test.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Incubation, Western Blot, Staining

    Protein ISGylation enhances type I IFN-mediated cytokine production. ( A ) Protein ISGylation promoted cytokine production in the presence of type I IFN treatment. Bone marrow-derived macrophages (BMDMs) from WT and Ube1L-KO mice were treated with type I IFN (500 U/mL) for 24 h and then with LPS (100 ng/mL) for 2 h. The expression of several cytokines was measured by RT-quantitative PCR (qPCR). The mRNA levels in WT cells without any treatment were set as 1, and the relative mRNA levels in other conditions were normalized accordingly. Results are shown as mean + SEM from three independent experiments. P , student’s t -test. ( B ) TNF-α expression in BMDMs in the presence of different inhibitors. BMDMs from WT and Ube1L-KO mice were primed with type I IFN at time 0 and were treated with the chemical inhibitors MG132 (0.5 μM), NAC (20 mM), or SB203580 (40 μM), respectively, for 1 h beginning 23 h after IFN treatment. Then the cells were stimulated by LPS (100 ng/mL) for another 2 h. Expression of TNF-α was measured by RT-qPCR. Data are shown as mean ± SD ( n = 2 or 3). P , student’s t -test. ( C ) Immunofluorescence staining for FK1 (red), F4/80 (green), and p-p38 (red) in mouse colon tissue treated with 2% (wt/vol) DSS. Representative images from one of four or five mouse samples are shown. (Original magnification, 40×.)
    Figure Legend Snippet: Protein ISGylation enhances type I IFN-mediated cytokine production. ( A ) Protein ISGylation promoted cytokine production in the presence of type I IFN treatment. Bone marrow-derived macrophages (BMDMs) from WT and Ube1L-KO mice were treated with type I IFN (500 U/mL) for 24 h and then with LPS (100 ng/mL) for 2 h. The expression of several cytokines was measured by RT-quantitative PCR (qPCR). The mRNA levels in WT cells without any treatment were set as 1, and the relative mRNA levels in other conditions were normalized accordingly. Results are shown as mean + SEM from three independent experiments. P , student’s t -test. ( B ) TNF-α expression in BMDMs in the presence of different inhibitors. BMDMs from WT and Ube1L-KO mice were primed with type I IFN at time 0 and were treated with the chemical inhibitors MG132 (0.5 μM), NAC (20 mM), or SB203580 (40 μM), respectively, for 1 h beginning 23 h after IFN treatment. Then the cells were stimulated by LPS (100 ng/mL) for another 2 h. Expression of TNF-α was measured by RT-qPCR. Data are shown as mean ± SD ( n = 2 or 3). P , student’s t -test. ( C ) Immunofluorescence staining for FK1 (red), F4/80 (green), and p-p38 (red) in mouse colon tissue treated with 2% (wt/vol) DSS. Representative images from one of four or five mouse samples are shown. (Original magnification, 40×.)

    Techniques Used: Derivative Assay, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Immunofluorescence, Staining

    16) Product Images from "Prolactin and Estrogen Enhance the Activity of Activating Protein 1 in Breast Cancer Cells: Role of Extracellularly Regulated Kinase 1/2-Mediated Signals to c-fos"

    Article Title: Prolactin and Estrogen Enhance the Activity of Activating Protein 1 in Breast Cancer Cells: Role of Extracellularly Regulated Kinase 1/2-Mediated Signals to c-fos

    Journal: Molecular endocrinology (Baltimore, Md.)

    doi: 10.1210/me.2004-0339

    Role for p38 in PRL and E2 Activation of AP-1
    Figure Legend Snippet: Role for p38 in PRL and E2 Activation of AP-1

    Techniques Used: Activation Assay

    17) Product Images from "Ca2+ and CACNA1H mediate targeted suppression of breast cancer brain metastasis by AM RF EMF"

    Article Title: Ca2+ and CACNA1H mediate targeted suppression of breast cancer brain metastasis by AM RF EMF

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2019.05.038

    The inhibitory effect of BCF is mediated through Ca v 3.2 T-type channel and CAMKII/p38 MAPK pathway. (A) 231BrM and SKBrM3 cells were treated with Sham or random frequencies (RCF) or BCF for 7 days followed by thymidine incorporation cell proliferatin assay ( n = 5/group). (B) SKBrM3 cells were treated with 1 h, 3 h and 6 h-per day for 7 days and cell proliferation was examined at day 7 by thymidine incorporation assay. (C) Various cell lines were seeded on 96-well plates at day 7 after Sham or BCF treatment, and cell proliferation was examined at day 1, 3 and 5 by MTS assay ( n = 8/group). (D) 231BrM and SKBrM3 cells were exposed to Sham or BCF daily for 7 days in the presence of ethosuximide or vehicle, and cell proliferation was quantified by thymidine incorporation at day 7 ( n = 5/group). Results are normalised to Sham group. ( E -F) The T-type voltage-gated calcium channel subunit genes, Ca v 3.1, Ca v 3.2 or Ca v 3.3, were knocked down in 231BrM or SKBrM3 cells by shRNAs, and they were treated with BCF for 7 days followed by quantifying cell proliferation at day 7. (G) SKBrM3shCa v 3.2 cells were intracardially injected to NOD/SCID mice. At Day 30, ex vivo tumour signal in brain was quantified by bioluminescence. Right panel shows representative brain images. (H) Cells treated with Sham or BCF were stained with Fluo-4 calcium dye and the level of intracellular calcium level was examined using flow cytometry. The representative histogram is shown. (I) SKBrM3 cells treated with Sham or BCF in the presence of vehicle or ethosuximide were stained with Fluo-4-am dye and cytoplasmic calcium level was quantified by flow cytometry ( n = 6/group). (J) SKBrM3 cells cultured in media with or without calcium and examined for cytoplasmic calcium as in I. (K) The expression of total and activated p38 and phosphorylated JNK was examined in 231BrM cells treated with Sham or BCF for 7 days by western blot. α-tubulin was used as a loading control. (L) 231-BrM cells were treated with Sham or BCF or BCF in the presence of KN93 (5 μM) and protein levels of phospho-CAMKII, total CAMKII, phopho-p38 and total p38 were examined by western blot. (M) SKBrM3 cells treated with Sham or BCF for 7 days were subjected to cell cycle analysis using FACS. % of population in G1, S and G2 phase are shown (n = 5/group). (N) Enrichment of biocarta P38MAPK signature in breast cancer patients with or without brain metastasis incidence was analysed by GSEA. (O) p38WT or p38 D176A (active mutation) gene or vector control was ectopically expressed in SKBrM3 cells using retrovirus and expression, and cells were seeded on 96-well plate ( n = 500/well) and subjected to cell proliferation assay using MTS reagent at Day 5 ( n = 8/group). *, P -value
    Figure Legend Snippet: The inhibitory effect of BCF is mediated through Ca v 3.2 T-type channel and CAMKII/p38 MAPK pathway. (A) 231BrM and SKBrM3 cells were treated with Sham or random frequencies (RCF) or BCF for 7 days followed by thymidine incorporation cell proliferatin assay ( n = 5/group). (B) SKBrM3 cells were treated with 1 h, 3 h and 6 h-per day for 7 days and cell proliferation was examined at day 7 by thymidine incorporation assay. (C) Various cell lines were seeded on 96-well plates at day 7 after Sham or BCF treatment, and cell proliferation was examined at day 1, 3 and 5 by MTS assay ( n = 8/group). (D) 231BrM and SKBrM3 cells were exposed to Sham or BCF daily for 7 days in the presence of ethosuximide or vehicle, and cell proliferation was quantified by thymidine incorporation at day 7 ( n = 5/group). Results are normalised to Sham group. ( E -F) The T-type voltage-gated calcium channel subunit genes, Ca v 3.1, Ca v 3.2 or Ca v 3.3, were knocked down in 231BrM or SKBrM3 cells by shRNAs, and they were treated with BCF for 7 days followed by quantifying cell proliferation at day 7. (G) SKBrM3shCa v 3.2 cells were intracardially injected to NOD/SCID mice. At Day 30, ex vivo tumour signal in brain was quantified by bioluminescence. Right panel shows representative brain images. (H) Cells treated with Sham or BCF were stained with Fluo-4 calcium dye and the level of intracellular calcium level was examined using flow cytometry. The representative histogram is shown. (I) SKBrM3 cells treated with Sham or BCF in the presence of vehicle or ethosuximide were stained with Fluo-4-am dye and cytoplasmic calcium level was quantified by flow cytometry ( n = 6/group). (J) SKBrM3 cells cultured in media with or without calcium and examined for cytoplasmic calcium as in I. (K) The expression of total and activated p38 and phosphorylated JNK was examined in 231BrM cells treated with Sham or BCF for 7 days by western blot. α-tubulin was used as a loading control. (L) 231-BrM cells were treated with Sham or BCF or BCF in the presence of KN93 (5 μM) and protein levels of phospho-CAMKII, total CAMKII, phopho-p38 and total p38 were examined by western blot. (M) SKBrM3 cells treated with Sham or BCF for 7 days were subjected to cell cycle analysis using FACS. % of population in G1, S and G2 phase are shown (n = 5/group). (N) Enrichment of biocarta P38MAPK signature in breast cancer patients with or without brain metastasis incidence was analysed by GSEA. (O) p38WT or p38 D176A (active mutation) gene or vector control was ectopically expressed in SKBrM3 cells using retrovirus and expression, and cells were seeded on 96-well plate ( n = 500/well) and subjected to cell proliferation assay using MTS reagent at Day 5 ( n = 8/group). *, P -value

    Techniques Used: Thymidine Incorporation Assay, MTS Assay, Injection, Mouse Assay, Ex Vivo, Staining, Flow Cytometry, Cytometry, Cell Culture, Expressing, Western Blot, Cell Cycle Assay, FACS, Mutagenesis, Plasmid Preparation, Proliferation Assay

    18) Product Images from "HEV ORF3 downregulates TLR7 to inhibit the generation of type I interferon via impairment of multiple signaling pathways"

    Article Title: HEV ORF3 downregulates TLR7 to inhibit the generation of type I interferon via impairment of multiple signaling pathways

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-26975-4

    HEV ORF3 impairs the activation of NFκB, JAK/STAT and JNK/MAPK pathways. ( A ) HEV ORF3 strengthen p-ERK and inhibits p-P65, p-STAT1 and p-JNK, but does not affect p-P38. Lane 1: LO2 cells. Lane 2: LO2 cells transfected with pcDNA3.1-GFP. Lane 3: LO2 cells transfected with pcDNA3.1-GFP-ORF3. ( B .
    Figure Legend Snippet: HEV ORF3 impairs the activation of NFκB, JAK/STAT and JNK/MAPK pathways. ( A ) HEV ORF3 strengthen p-ERK and inhibits p-P65, p-STAT1 and p-JNK, but does not affect p-P38. Lane 1: LO2 cells. Lane 2: LO2 cells transfected with pcDNA3.1-GFP. Lane 3: LO2 cells transfected with pcDNA3.1-GFP-ORF3. ( B .

    Techniques Used: Activation Assay, Transfection

    19) Product Images from "Beneficial Effects of Deoxyshikonin on Delayed Wound Healing in Diabetic Mice"

    Article Title: Beneficial Effects of Deoxyshikonin on Delayed Wound Healing in Diabetic Mice

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19113660

    Effect of p38 and extracellular signal-regulated kinase (ERK) on deoxyshikonin-promoted tube formation in HUVECs. ( A ) HUVECs were pretreated with inhibitors (SB203580 or U0126) then treated deoxyshikonin for 24 h. After that, the amount of expressed protein of each antibody (phosphorylated-p38, p38, phosphorylated-ERK, and ERK) was quantified. Western blot was performed three times using an independently prepared cell lysate. ( B ) The relative intensity of Western blot bands was measured using the ImageJ software. ( C ) Photographs of capillary-like tube formation in HUVECs (treated with inhibitors) on Matrigel after incubation with deoxyshikonin 3 μM after 24 h. ( D ) The relative lengths of tubes were measured using the ImageJ software. Data are expressed as means ± SEM. Similar results were obtained from three independent experiments; * p
    Figure Legend Snippet: Effect of p38 and extracellular signal-regulated kinase (ERK) on deoxyshikonin-promoted tube formation in HUVECs. ( A ) HUVECs were pretreated with inhibitors (SB203580 or U0126) then treated deoxyshikonin for 24 h. After that, the amount of expressed protein of each antibody (phosphorylated-p38, p38, phosphorylated-ERK, and ERK) was quantified. Western blot was performed three times using an independently prepared cell lysate. ( B ) The relative intensity of Western blot bands was measured using the ImageJ software. ( C ) Photographs of capillary-like tube formation in HUVECs (treated with inhibitors) on Matrigel after incubation with deoxyshikonin 3 μM after 24 h. ( D ) The relative lengths of tubes were measured using the ImageJ software. Data are expressed as means ± SEM. Similar results were obtained from three independent experiments; * p

    Techniques Used: Western Blot, Software, Incubation

    20) Product Images from "Extracellular ADP facilitates monocyte recruitment in bacterial infection via ERK signaling"

    Article Title: Extracellular ADP facilitates monocyte recruitment in bacterial infection via ERK signaling

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/cmi.2016.56

    ADP activates cAMP-extracellular signal-regulated kinase (ERK1)2 signaling. ( a ) RAW 264.7 cells and ( b ) BMDMs (only for p-ERK) were stimulated with ADP, and samples from a time course were subjected to western blot analysis to detect the phosphorylation of ERK1/2, JNK and p38 MAPK. ( c ) In addition, the phosphorylation of PKA was measured as described above. RAW 264.7 cells were incubated with or without ( d ) an ERK1/2 inhibitor (U0126, 10 μM), an adenylyl cyclase activator (forskolin, 100 μ) or ( e ) a cAMP-dependent PKA activator (8-Bromo-cAMP, 50 μM) for 30 min before stimulation with 100 μM ADP. Two hours later, the cell lysates were subjected to western blot analysis to detect p-ERK. Semi-quantitative analysis and representative figures are shown. The immunoblot results are representative of at least three independent experiments. The results are expressed as the mean±s.e.m., and statistical analysis was performed with one-way ANOVA; * P
    Figure Legend Snippet: ADP activates cAMP-extracellular signal-regulated kinase (ERK1)2 signaling. ( a ) RAW 264.7 cells and ( b ) BMDMs (only for p-ERK) were stimulated with ADP, and samples from a time course were subjected to western blot analysis to detect the phosphorylation of ERK1/2, JNK and p38 MAPK. ( c ) In addition, the phosphorylation of PKA was measured as described above. RAW 264.7 cells were incubated with or without ( d ) an ERK1/2 inhibitor (U0126, 10 μM), an adenylyl cyclase activator (forskolin, 100 μ) or ( e ) a cAMP-dependent PKA activator (8-Bromo-cAMP, 50 μM) for 30 min before stimulation with 100 μM ADP. Two hours later, the cell lysates were subjected to western blot analysis to detect p-ERK. Semi-quantitative analysis and representative figures are shown. The immunoblot results are representative of at least three independent experiments. The results are expressed as the mean±s.e.m., and statistical analysis was performed with one-way ANOVA; * P

    Techniques Used: Western Blot, Incubation

    21) Product Images from "A standardized herbal extract PM014 ameliorates pulmonary fibrosis by suppressing the TGF-β1 pathway"

    Article Title: A standardized herbal extract PM014 ameliorates pulmonary fibrosis by suppressing the TGF-β1 pathway

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-35320-8

    PM014 suppresses TGF-β1 pathway through inactivating canonical and non-canonical signaling in A549 cells. Human epithelial alveolar A549 cells were stimulated with TGF-β1 (20 ng/ml) and PM014 (0.1, 0.4, 1.0 mg/ml) for 48 h. ( a ) Western blotting analysis of phosphorylation levels of Smad2 and Smad3 and p38 MAPK. ( b ) Densitometry of the immune blot was analyzed quantitatively. Expression of p-Smad2/3 and p-p38 is represented as a percentage compared to that of the control DMSO-treated group. ( c – e ) Real-time qPCR was performed to examine the mRNA levels of Slug, Snail, and TGF-β1. The cropped gels are used in the figure, and uncropped gels are presented in Fig. S4. Data are presented as mean ± SE, n = 3–4, * P
    Figure Legend Snippet: PM014 suppresses TGF-β1 pathway through inactivating canonical and non-canonical signaling in A549 cells. Human epithelial alveolar A549 cells were stimulated with TGF-β1 (20 ng/ml) and PM014 (0.1, 0.4, 1.0 mg/ml) for 48 h. ( a ) Western blotting analysis of phosphorylation levels of Smad2 and Smad3 and p38 MAPK. ( b ) Densitometry of the immune blot was analyzed quantitatively. Expression of p-Smad2/3 and p-p38 is represented as a percentage compared to that of the control DMSO-treated group. ( c – e ) Real-time qPCR was performed to examine the mRNA levels of Slug, Snail, and TGF-β1. The cropped gels are used in the figure, and uncropped gels are presented in Fig. S4. Data are presented as mean ± SE, n = 3–4, * P

    Techniques Used: Western Blot, Expressing, Real-time Polymerase Chain Reaction

    22) Product Images from "Hydroxysafflor Yellow A Suppresses Platelet Activating Factor-Induced Activation of Human Small Airway Epithelial Cells"

    Article Title: Hydroxysafflor Yellow A Suppresses Platelet Activating Factor-Induced Activation of Human Small Airway Epithelial Cells

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2018.00859

    Effects of HSYA on MAPK (ERK, JNK, p38) pathway and transcriptional activities of NFκB and AP1 in PAF-stimulated HSAECs. Phosphorylation levels of ERK (A) , JNK (C) , p38 (E) , and IκB (G) were determined by western blot and quantified by densitometric analysis (B, D, F, H) . Binding activities of NFκB (I) and AP1 (J) were determined by dual-luciferase assay. Data presented as mean ± SD, n = 3 per group. ANOVA, p
    Figure Legend Snippet: Effects of HSYA on MAPK (ERK, JNK, p38) pathway and transcriptional activities of NFκB and AP1 in PAF-stimulated HSAECs. Phosphorylation levels of ERK (A) , JNK (C) , p38 (E) , and IκB (G) were determined by western blot and quantified by densitometric analysis (B, D, F, H) . Binding activities of NFκB (I) and AP1 (J) were determined by dual-luciferase assay. Data presented as mean ± SD, n = 3 per group. ANOVA, p

    Techniques Used: Western Blot, Binding Assay, Luciferase

    23) Product Images from "Triptolide induces Sertoli cell apoptosis in mice via ROS/JNK-dependent activation of the mitochondrial pathway and inhibition of Nrf2-mediated antioxidant response"

    Article Title: Triptolide induces Sertoli cell apoptosis in mice via ROS/JNK-dependent activation of the mitochondrial pathway and inhibition of Nrf2-mediated antioxidant response

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2017.95

    Activation of JNK phosphorylation during TP-induced apoptosis in cultured TM4 cells. (A) Cells were cultured in 6-well plates until confluent, and the medium was replaced with 1% serum medium in the presence or absence of TP (125, 250 and 500 nmol/L) for 24 and 48 h. The cells were lysed and MAPKs proteins were analyzed Western blot. (B) Densitometry scanning analysis of ratio of p-JNK/total JNK, p-p38/total p38 and p-ERK1/2/total ERK1/2. Data represent the mean±SD of three independent experiments. * P
    Figure Legend Snippet: Activation of JNK phosphorylation during TP-induced apoptosis in cultured TM4 cells. (A) Cells were cultured in 6-well plates until confluent, and the medium was replaced with 1% serum medium in the presence or absence of TP (125, 250 and 500 nmol/L) for 24 and 48 h. The cells were lysed and MAPKs proteins were analyzed Western blot. (B) Densitometry scanning analysis of ratio of p-JNK/total JNK, p-p38/total p38 and p-ERK1/2/total ERK1/2. Data represent the mean±SD of three independent experiments. * P

    Techniques Used: Activation Assay, Cell Culture, Western Blot

    24) Product Images from "Overexpression of CD44 Standard Isoform Upregulates HIF-1α Signaling in Hypoxic Breast Cancer Cells"

    Article Title: Overexpression of CD44 Standard Isoform Upregulates HIF-1α Signaling in Hypoxic Breast Cancer Cells

    Journal: Biomolecules & Therapeutics

    doi: 10.4062/biomolther.2018.116

    ERK signaling is involved in the CD44s-mediated HIF-1α accumulation. (A) Protein levels of total/phosphorylated ERK (p-ERK), total/phosphorylated p38 (p-p38), total/phosphorylated JNK (p-JNK), and total/phosphorylated AKT (p-AKT) were determined in pBabe-MCF7 and pCD44s-MCF7 cells after incubation in normoxic or hypoxic condition. Quantitative protein levels are represented as mean ± SE from three experiments. a p
    Figure Legend Snippet: ERK signaling is involved in the CD44s-mediated HIF-1α accumulation. (A) Protein levels of total/phosphorylated ERK (p-ERK), total/phosphorylated p38 (p-p38), total/phosphorylated JNK (p-JNK), and total/phosphorylated AKT (p-AKT) were determined in pBabe-MCF7 and pCD44s-MCF7 cells after incubation in normoxic or hypoxic condition. Quantitative protein levels are represented as mean ± SE from three experiments. a p

    Techniques Used: Incubation

    25) Product Images from "Peiminine Protects against Lipopolysaccharide-Induced Mastitis by Inhibiting the AKT/NF-κB, ERK1/2 and p38 Signaling Pathways"

    Article Title: Peiminine Protects against Lipopolysaccharide-Induced Mastitis by Inhibiting the AKT/NF-κB, ERK1/2 and p38 Signaling Pathways

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19092637

    Peiminine inhibits the phosphorylation of the AKT, NF-κB p65, ERK1/2, and p38 signaling pathways in mMECs. The results for the protein levels of ( A , B ) p-AKT, ( A , C ) p-NF-κB p65, ( A , D ) p-ERK1/2, and ( A , E ) p-p38 were measured by western blotting. The data are presented as the mean ± SD ( n = 6). The number sign (#) indicates a significant difference from the NT group at p
    Figure Legend Snippet: Peiminine inhibits the phosphorylation of the AKT, NF-κB p65, ERK1/2, and p38 signaling pathways in mMECs. The results for the protein levels of ( A , B ) p-AKT, ( A , C ) p-NF-κB p65, ( A , D ) p-ERK1/2, and ( A , E ) p-p38 were measured by western blotting. The data are presented as the mean ± SD ( n = 6). The number sign (#) indicates a significant difference from the NT group at p

    Techniques Used: Western Blot

    Peiminine inhibits LPS-induced inflammation by suppressing the phosphorylation of the AKT, NF-κB p65, ERK1/2, and p38 signaling pathways.
    Figure Legend Snippet: Peiminine inhibits LPS-induced inflammation by suppressing the phosphorylation of the AKT, NF-κB p65, ERK1/2, and p38 signaling pathways.

    Techniques Used:

    Peiminine inhibits the phosphorylation of the AKT, nuclear factor-κB (NF-κB) p65, ERK1/2 and p38 signaling pathways in the mammary tissues. The results for the protein levels of ( A , B ) p-AKT, ( A , C ) p-NF-κB p65, ( A , D ) p-ERK1/2, and ( A , E ) p-p38 were measured by western blotting. The data are presented as the mean ± SD ( n = 6). The number sign (#) indicates a significant difference from the NT group at p
    Figure Legend Snippet: Peiminine inhibits the phosphorylation of the AKT, nuclear factor-κB (NF-κB) p65, ERK1/2 and p38 signaling pathways in the mammary tissues. The results for the protein levels of ( A , B ) p-AKT, ( A , C ) p-NF-κB p65, ( A , D ) p-ERK1/2, and ( A , E ) p-p38 were measured by western blotting. The data are presented as the mean ± SD ( n = 6). The number sign (#) indicates a significant difference from the NT group at p

    Techniques Used: Western Blot

    26) Product Images from "Pilloin, A Flavonoid Isolated from Aquilaria sinensis, Exhibits Anti-Inflammatory Activity In Vitro and In Vivo"

    Article Title: Pilloin, A Flavonoid Isolated from Aquilaria sinensis, Exhibits Anti-Inflammatory Activity In Vitro and In Vivo

    Journal: Molecules

    doi: 10.3390/molecules23123177

    Effects of pilloin on MAPK signalling pathways in LPS-stimulated RAW 264.7 macrophages. RAW 264.7 cells (10 6 cells in MP-6 plates) were treated with vehicle, pilloin or the indicated inhibitor for 6 h before incubation with LPS (100 ng/mL) for 20 min. Cell lysates were analysed by Western blot to detect the activation of ( A ) JNK, ( B ) ERK and ( C ) p38. PD98059 (ERK inhibitor), SB203580 (p38 inhibitor) and SP600125 (JNK inhibitor) served as positive controls. * indicates a significant difference versus the LPS-treated group ( p
    Figure Legend Snippet: Effects of pilloin on MAPK signalling pathways in LPS-stimulated RAW 264.7 macrophages. RAW 264.7 cells (10 6 cells in MP-6 plates) were treated with vehicle, pilloin or the indicated inhibitor for 6 h before incubation with LPS (100 ng/mL) for 20 min. Cell lysates were analysed by Western blot to detect the activation of ( A ) JNK, ( B ) ERK and ( C ) p38. PD98059 (ERK inhibitor), SB203580 (p38 inhibitor) and SP600125 (JNK inhibitor) served as positive controls. * indicates a significant difference versus the LPS-treated group ( p

    Techniques Used: Incubation, Western Blot, Activation Assay

    27) Product Images from "Magnolol inhibits growth and induces apoptosis in esophagus cancer KYSE-150 cell lines via the MAP kinase pathway"

    Article Title: Magnolol inhibits growth and induces apoptosis in esophagus cancer KYSE-150 cell lines via the MAP kinase pathway

    Journal: Journal of Thoracic Disease

    doi: 10.21037/jtd.2019.07.46

    Magnolol induces activation of the MAP Kinase pathway. KYSE-150 cells were treated with different concentrations of magnolol and then cells were analyzed for phosphorylation of ERK, JNK and p38. Phosphorylation of ERK was significantly increased after magnolol treatment.
    Figure Legend Snippet: Magnolol induces activation of the MAP Kinase pathway. KYSE-150 cells were treated with different concentrations of magnolol and then cells were analyzed for phosphorylation of ERK, JNK and p38. Phosphorylation of ERK was significantly increased after magnolol treatment.

    Techniques Used: Activation Assay

    28) Product Images from "Cadmium Induces Acute Liver Injury by Inhibiting Nrf2 and the Role of NF-κB, NLRP3, and MAPKs Signaling Pathway"

    Article Title: Cadmium Induces Acute Liver Injury by Inhibiting Nrf2 and the Role of NF-κB, NLRP3, and MAPKs Signaling Pathway

    Journal: International Journal of Environmental Research and Public Health

    doi: 10.3390/ijerph17010138

    Cd activated nuclear factor-κB (NF-κB), Nod-like receptor 3 (NLRP3), and mitogen-activated protein kinases (MAPKs) in mouse liver ( n = 4). Phosphorylation of p65, ERK, JNK and p38 and the expression of NLRP3 were evaluated by Western blotting. * p
    Figure Legend Snippet: Cd activated nuclear factor-κB (NF-κB), Nod-like receptor 3 (NLRP3), and mitogen-activated protein kinases (MAPKs) in mouse liver ( n = 4). Phosphorylation of p65, ERK, JNK and p38 and the expression of NLRP3 were evaluated by Western blotting. * p

    Techniques Used: Expressing, Western Blot

    29) Product Images from "Curcumin suppresses transforming growth factor-β1-induced cardiac fibroblast differentiation via inhibition of Smad-2 and p38 MAPK signaling pathways"

    Article Title: Curcumin suppresses transforming growth factor-β1-induced cardiac fibroblast differentiation via inhibition of Smad-2 and p38 MAPK signaling pathways

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2016.2969

    Curcumin administration inhibited the transforming growth factor (TGF)-β1-induced activation of p38 in cardiac fibroblasts (CFs). CFs were pretreated with 20 µmol/l curcumin, 10 µM SB431542 (Smad2 inhibitor) or 10 µM SB203580
    Figure Legend Snippet: Curcumin administration inhibited the transforming growth factor (TGF)-β1-induced activation of p38 in cardiac fibroblasts (CFs). CFs were pretreated with 20 µmol/l curcumin, 10 µM SB431542 (Smad2 inhibitor) or 10 µM SB203580

    Techniques Used: Activation Assay

    Effects of transforming growth factor (TGF)-β1 on the phosphorylation of (A) Smad2 and (B) p38 over 60 min, as determined by western blotting. Total Smad2 and p38 was used as the control. Densitometric analysis was performed to quantify the phosphorylation
    Figure Legend Snippet: Effects of transforming growth factor (TGF)-β1 on the phosphorylation of (A) Smad2 and (B) p38 over 60 min, as determined by western blotting. Total Smad2 and p38 was used as the control. Densitometric analysis was performed to quantify the phosphorylation

    Techniques Used: Western Blot

    Smad2 and p38 inhibitors, SB431542 and SB20380 respectively, suppressed (A) α-smooth muscle actin (SMA) and (B) collagen I (ColI) expression levels. Cardiac fibroblasts (CFs) were treated with 10 ng/ml transforming growth factor (TGF)-β1
    Figure Legend Snippet: Smad2 and p38 inhibitors, SB431542 and SB20380 respectively, suppressed (A) α-smooth muscle actin (SMA) and (B) collagen I (ColI) expression levels. Cardiac fibroblasts (CFs) were treated with 10 ng/ml transforming growth factor (TGF)-β1

    Techniques Used: Expressing

    30) Product Images from "Peroxiredoxin-1 Overexpression Attenuates Doxorubicin-Induced Cardiotoxicity by Inhibiting Oxidative Stress and Cardiomyocyte Apoptosis"

    Article Title: Peroxiredoxin-1 Overexpression Attenuates Doxorubicin-Induced Cardiotoxicity by Inhibiting Oxidative Stress and Cardiomyocyte Apoptosis

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2020/2405135

    Prdx1 overexpression inhibits ASK1/p38 pathway activation. (a) Western blots showing the p-ASK1, ASK1, p-p38, and p38 levels in the hearts of mice ( n = 4). (b) Western blots showing the p-ASK1, ASK1, p-p38, and p38 levels in NVRMs ( n = 4). ∗ P
    Figure Legend Snippet: Prdx1 overexpression inhibits ASK1/p38 pathway activation. (a) Western blots showing the p-ASK1, ASK1, p-p38, and p38 levels in the hearts of mice ( n = 4). (b) Western blots showing the p-ASK1, ASK1, p-p38, and p38 levels in NVRMs ( n = 4). ∗ P

    Techniques Used: Over Expression, Activation Assay, Western Blot, Mouse Assay

    31) Product Images from "Heparanase Released From Mesenchymal Stem Cells Activates Integrin beta1/HIF-2alpha/Flk-1 Signaling and Promotes Endothelial Cell Migration and Angiogenesis"

    Article Title: Heparanase Released From Mesenchymal Stem Cells Activates Integrin beta1/HIF-2alpha/Flk-1 Signaling and Promotes Endothelial Cell Migration and Angiogenesis

    Journal: Stem cells (Dayton, Ohio)

    doi: 10.1002/stem.1995

    Enhanced cell migration by MSC hpa via Flk-1/p38. (A) : Representative images showing cell migration of HUVECs after cocultured with conditioned medium derived from MSC WT , MSC null , MSC hpa , MSC hpa-KD , respectively; Bar = 50 μm. (B, C) : Bar graph showing quantitative analysis of cell migration of HUVECs in each group; n = 3, * denotes p
    Figure Legend Snippet: Enhanced cell migration by MSC hpa via Flk-1/p38. (A) : Representative images showing cell migration of HUVECs after cocultured with conditioned medium derived from MSC WT , MSC null , MSC hpa , MSC hpa-KD , respectively; Bar = 50 μm. (B, C) : Bar graph showing quantitative analysis of cell migration of HUVECs in each group; n = 3, * denotes p

    Techniques Used: Migration, Derivative Assay

    32) Product Images from "Cardiac Tissue Engineering in Magnetically Actuated Scaffolds"

    Article Title: Cardiac Tissue Engineering in Magnetically Actuated Scaffolds

    Journal: Nanotechnology

    doi: 10.1088/0957-4484/25/1/014009

    Cardiac cells adhesion onto the macroporous MNP-impregnated scaffolds. (A) Immunostaining for vinculin (green), F-actin (red) and nuclei (blue), 1, 3 and 24 h post-seeding. Magnification is ×20. AKT (B) and P38 (C) phosphorylation by western blot
    Figure Legend Snippet: Cardiac cells adhesion onto the macroporous MNP-impregnated scaffolds. (A) Immunostaining for vinculin (green), F-actin (red) and nuclei (blue), 1, 3 and 24 h post-seeding. Magnification is ×20. AKT (B) and P38 (C) phosphorylation by western blot

    Techniques Used: Immunostaining, Western Blot

    33) Product Images from "Bacillus subtilis Fermentation of Malva verticillata Leaves Enhances Antioxidant Activity and Osteoblast Differentiation"

    Article Title: Bacillus subtilis Fermentation of Malva verticillata Leaves Enhances Antioxidant Activity and Osteoblast Differentiation

    Journal: Foods

    doi: 10.3390/foods9050671

    Western blot showing activation of transforming growth factor-β (TGF-β) signaling pathway by MVB treatment in C3H10T1/2 cells. ( A ) Time-course analysis of the non-canonical signaling of TGF-β pathway after treatment with 100 μg/mL MVB. ( B ) Time-course analysis of Runx2 and Osterix after treatment with 100 μg/mL MVB. ( C ) Pre-treating cells with inhibitors of TGF-β signaling pathway attenuated MVB (100 μg/mL)-induced p38 and ERK activation. Significant difference compared with control: * p
    Figure Legend Snippet: Western blot showing activation of transforming growth factor-β (TGF-β) signaling pathway by MVB treatment in C3H10T1/2 cells. ( A ) Time-course analysis of the non-canonical signaling of TGF-β pathway after treatment with 100 μg/mL MVB. ( B ) Time-course analysis of Runx2 and Osterix after treatment with 100 μg/mL MVB. ( C ) Pre-treating cells with inhibitors of TGF-β signaling pathway attenuated MVB (100 μg/mL)-induced p38 and ERK activation. Significant difference compared with control: * p

    Techniques Used: Western Blot, Activation Assay

    34) Product Images from "P38/MAPK contributes to endothelial barrier dysfunction via MAP4 phosphorylation-dependent microtubule disassembly in inflammation-induced acute lung injury"

    Article Title: P38/MAPK contributes to endothelial barrier dysfunction via MAP4 phosphorylation-dependent microtubule disassembly in inflammation-induced acute lung injury

    Journal: Scientific Reports

    doi: 10.1038/srep08895

    P38/MAPK activation mediates MAP4 phosphorylation in inflammation-induced ALI. (a) Western blotting was used to detect phospho-p38 (p-P38) and P38 following treatment with LPS or TNF-α (500 ng/ml for 1, 3, 6, and 12 hr). *P
    Figure Legend Snippet: P38/MAPK activation mediates MAP4 phosphorylation in inflammation-induced ALI. (a) Western blotting was used to detect phospho-p38 (p-P38) and P38 following treatment with LPS or TNF-α (500 ng/ml for 1, 3, 6, and 12 hr). *P

    Techniques Used: Activation Assay, Western Blot

    Role of p38/MAPK activation in LPS- and TNF-α-induced endothelial barrier dysfunction and MT disassembly. (a) Cells were pretreated with SB203580 (5 μM) for 1 hr, and CMV-null or MKK6 (Glu) was transfected into HPMECs for 72 hr before the LPS or TNF-α (500 ng/ml) treatment. The permeability of endothelial cells was assessed by measuring the influx of FITC-conjugated dextran and the TER across the cells. The data are represented as the mean ± SEM (n = 3). *P
    Figure Legend Snippet: Role of p38/MAPK activation in LPS- and TNF-α-induced endothelial barrier dysfunction and MT disassembly. (a) Cells were pretreated with SB203580 (5 μM) for 1 hr, and CMV-null or MKK6 (Glu) was transfected into HPMECs for 72 hr before the LPS or TNF-α (500 ng/ml) treatment. The permeability of endothelial cells was assessed by measuring the influx of FITC-conjugated dextran and the TER across the cells. The data are represented as the mean ± SEM (n = 3). *P

    Techniques Used: Activation Assay, Transfection, Permeability

    35) Product Images from "Glutathione S-transferase P protects against cyclophosphamide-induced cardiotoxicity in mice"

    Article Title: Glutathione S-transferase P protects against cyclophosphamide-induced cardiotoxicity in mice

    Journal: Toxicology and applied pharmacology

    doi: 10.1016/j.taap.2015.03.029

    Cyclophosphamide (CY)-induced phosphorylation of JNK, c-Jun, ERK1/2 and p38 in hearts of WT and GSTP-null mice
    Figure Legend Snippet: Cyclophosphamide (CY)-induced phosphorylation of JNK, c-Jun, ERK1/2 and p38 in hearts of WT and GSTP-null mice

    Techniques Used: Mouse Assay

    36) Product Images from "Selective control of type I IFN induction by the Rac activator DOCK2 during TLR-mediated plasmacytoid dendritic cell activation"

    Article Title: Selective control of type I IFN induction by the Rac activator DOCK2 during TLR-mediated plasmacytoid dendritic cell activation

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20091776

    DOCK2-mediated Rac activation is critical for IKK-α activation in pDCs. (A) Subcellular localization of IRF-7 was compared between WT and Dock2 −/− pDCs after stimulation with CpG-A. DAPI was used to stain nuclei. Representative images of three independent experiments are shown. Bar, 5 µm. (B) BM-derived pDCs from WT and Dock2 −/− mice were stimulated with 3 µM CpG-A for the indicated times and analyzed for phosphorylation of Akt, ERK, JNK, or p38. Data are representative of at least two independent experiments. (C and D) BM-derived WT and Dock2 −/− pDCs were stimulated with 3 µM CpG-A for the indicated times and analyzed for the expression (C) and autophosphorylation (D) of IRAK-1. Data are representative of two independent experiments. (E–G) BM-derived pDCs from WT, Dock2 −/− , and Tlr9 −/− mice were stimulated with 3 µM CpG-A for the indicated times and analyzed for serine phosphorylation of IKK-α at positions 176 and 180. (G) Before stimulation, cells were treated with bacterially expressed GFP-tagged Tat–T17N-Rac or Tat–WT Rac (500 nM each) for 30 min at 37°C. Data are representative of three (E and G) or two (F) independent experiments.
    Figure Legend Snippet: DOCK2-mediated Rac activation is critical for IKK-α activation in pDCs. (A) Subcellular localization of IRF-7 was compared between WT and Dock2 −/− pDCs after stimulation with CpG-A. DAPI was used to stain nuclei. Representative images of three independent experiments are shown. Bar, 5 µm. (B) BM-derived pDCs from WT and Dock2 −/− mice were stimulated with 3 µM CpG-A for the indicated times and analyzed for phosphorylation of Akt, ERK, JNK, or p38. Data are representative of at least two independent experiments. (C and D) BM-derived WT and Dock2 −/− pDCs were stimulated with 3 µM CpG-A for the indicated times and analyzed for the expression (C) and autophosphorylation (D) of IRAK-1. Data are representative of two independent experiments. (E–G) BM-derived pDCs from WT, Dock2 −/− , and Tlr9 −/− mice were stimulated with 3 µM CpG-A for the indicated times and analyzed for serine phosphorylation of IKK-α at positions 176 and 180. (G) Before stimulation, cells were treated with bacterially expressed GFP-tagged Tat–T17N-Rac or Tat–WT Rac (500 nM each) for 30 min at 37°C. Data are representative of three (E and G) or two (F) independent experiments.

    Techniques Used: Activation Assay, Staining, Derivative Assay, Mouse Assay, Expressing

    37) Product Images from "MAPK signaling pathways and HDAC3 activity are disrupted during differentiation of emerin-null myogenic progenitor cells"

    Article Title: MAPK signaling pathways and HDAC3 activity are disrupted during differentiation of emerin-null myogenic progenitor cells

    Journal: Disease Models & Mechanisms

    doi: 10.1242/dmm.028787

    p38 MAPK inhibition by SB203580 treatment prevents myogenic differentiation in wild-type and emerin-null myogenic progenitors. (A) Timeline showing the timing of SB203580 addition and sample collection for western blot analysis of whole cell lysates during differentiation. (B-I′) Representative images of vehicle-treated wild-type (B-E) or emerin-null (F-I) cells and SB203580-treated wild-type (B′-E′) or emerin-null (F′-I′) cells 36 h after differentiation induction. 40× magnification. (J-L) Quantification of > 1000 nuclei for each experimental treatment ( n ≥3) was done to determine the percentage of myogenic progenitors in the cell cycle (J), percentage of cells expressing MyHC (K) and the number of myotubes formed (L) 36 h post-differentiation induction. Results are mean±s.d. of n ≥3; N.S., not significant; * P
    Figure Legend Snippet: p38 MAPK inhibition by SB203580 treatment prevents myogenic differentiation in wild-type and emerin-null myogenic progenitors. (A) Timeline showing the timing of SB203580 addition and sample collection for western blot analysis of whole cell lysates during differentiation. (B-I′) Representative images of vehicle-treated wild-type (B-E) or emerin-null (F-I) cells and SB203580-treated wild-type (B′-E′) or emerin-null (F′-I′) cells 36 h after differentiation induction. 40× magnification. (J-L) Quantification of > 1000 nuclei for each experimental treatment ( n ≥3) was done to determine the percentage of myogenic progenitors in the cell cycle (J), percentage of cells expressing MyHC (K) and the number of myotubes formed (L) 36 h post-differentiation induction. Results are mean±s.d. of n ≥3; N.S., not significant; * P

    Techniques Used: Inhibition, Western Blot, Expressing

    ERK, p38 MAPK and HDAC3 regulate specific transition stages during myogenic differentiation. The stages of myogenic differentiation in wild-type (top) and emerin-null (bottom) myogenic progenitors are illustrated. Inhibition of p38 MAPK activity blocks cell cycle withdrawal and commitment to myogenic differentiation in both wild-type and emerin-null progenitors. HDAC3 inhibition blocks differentiation commitment and myotube formation in both wild-type and emerin-null progenitors. ERK inhibition rescues differentiation commitment and myotube formation in emerin-null progenitors with no effect on wild-type differentiation. Activation of HDAC3 catalytic activity rescues myotube formation in emerin-null myogenic progenitors with no effect on wild-type differentiation. Green arrows indicate rescue; red lines indicate blockade of differentiation progression.
    Figure Legend Snippet: ERK, p38 MAPK and HDAC3 regulate specific transition stages during myogenic differentiation. The stages of myogenic differentiation in wild-type (top) and emerin-null (bottom) myogenic progenitors are illustrated. Inhibition of p38 MAPK activity blocks cell cycle withdrawal and commitment to myogenic differentiation in both wild-type and emerin-null progenitors. HDAC3 inhibition blocks differentiation commitment and myotube formation in both wild-type and emerin-null progenitors. ERK inhibition rescues differentiation commitment and myotube formation in emerin-null progenitors with no effect on wild-type differentiation. Activation of HDAC3 catalytic activity rescues myotube formation in emerin-null myogenic progenitors with no effect on wild-type differentiation. Green arrows indicate rescue; red lines indicate blockade of differentiation progression.

    Techniques Used: Inhibition, Activity Assay, Activation Assay

    p38 MAPK phosphorylation is decreased by treatment with the p38 MAPK inhibitor SB203580 in differentiating myogenic progenitors. (A) Western blotting of whole cell lysates treated with SB203580 was performed to analyze activation of p38 MAPK during differentiation of wild-type or emerin-null progenitors. DMSO treatment was the control. Three biological replicates are shown for each treatment. (B) Densitometry was performed and phosphorylated p38 MAPK was normalized to total p38 MAPK protein in each sample. Levels of phosphorylated p38 MAPK for each condition were normalized to DMSO-treated wild-type cells. Results are mean±s.d. of n =3 for each condition; * P
    Figure Legend Snippet: p38 MAPK phosphorylation is decreased by treatment with the p38 MAPK inhibitor SB203580 in differentiating myogenic progenitors. (A) Western blotting of whole cell lysates treated with SB203580 was performed to analyze activation of p38 MAPK during differentiation of wild-type or emerin-null progenitors. DMSO treatment was the control. Three biological replicates are shown for each treatment. (B) Densitometry was performed and phosphorylated p38 MAPK was normalized to total p38 MAPK protein in each sample. Levels of phosphorylated p38 MAPK for each condition were normalized to DMSO-treated wild-type cells. Results are mean±s.d. of n =3 for each condition; * P

    Techniques Used: Western Blot, Activation Assay

    38) Product Images from "Paclitaxel induces apoptosis and reduces proliferation by targeting epidermal growth factor receptor signaling pathway in oral cavity squamous cell carcinoma"

    Article Title: Paclitaxel induces apoptosis and reduces proliferation by targeting epidermal growth factor receptor signaling pathway in oral cavity squamous cell carcinoma

    Journal: Oncology Letters

    doi: 10.3892/ol.2015.3499

    EGFR signaling pathway was significantly suppressed with PTX treatment. (A) Histochemistry staining demonstrated that EGFR was significantly increased in human oral cancer tissues compared with the normal tissue. (B) Western blot analysis revealed that PTX inhibited EGF-induced EGFR activation. (C) Western blot analysis demonstrated reduced c-Jun N-terminal kinases, extracellular signal-regulated kinases, p38 activation of EGFR signaling pathways. Data represent the means ± standard error of the mean, n=3 independent experiments. *P
    Figure Legend Snippet: EGFR signaling pathway was significantly suppressed with PTX treatment. (A) Histochemistry staining demonstrated that EGFR was significantly increased in human oral cancer tissues compared with the normal tissue. (B) Western blot analysis revealed that PTX inhibited EGF-induced EGFR activation. (C) Western blot analysis demonstrated reduced c-Jun N-terminal kinases, extracellular signal-regulated kinases, p38 activation of EGFR signaling pathways. Data represent the means ± standard error of the mean, n=3 independent experiments. *P

    Techniques Used: Staining, Western Blot, Activation Assay

    39) Product Images from "PKD PREVENTS H2O2-INDUCED APOPTOSIS VIA NF-?B AND p38 MAPK"

    Article Title: PKD PREVENTS H2O2-INDUCED APOPTOSIS VIA NF-?B AND p38 MAPK

    Journal:

    doi: 10.1016/j.bbrc.2008.11.106

    PKD1 affects H 2 O 2 -induced p38 MAPK phosphorylation but not ERK1/2 and JNK >
    Figure Legend Snippet: PKD1 affects H 2 O 2 -induced p38 MAPK phosphorylation but not ERK1/2 and JNK >

    Techniques Used:

    Phosphorylation of p38 MAPK induced by H 2 O 2 mediates through MKKs
    Figure Legend Snippet: Phosphorylation of p38 MAPK induced by H 2 O 2 mediates through MKKs

    Techniques Used:

    40) Product Images from "Phospholipid transfer protein and alpha-1 antitrypsin regulate Hck kinase activity during neutrophil degranulation"

    Article Title: Phospholipid transfer protein and alpha-1 antitrypsin regulate Hck kinase activity during neutrophil degranulation

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-33851-8

    Neutrophils from Pltp deficient mice has elevated tyrosine kinase activity of Hck. Neutrophils were collected from C57BL/6 and Pltp −/− mice. Neutrophils were exposed to AAT prior to fMLP at 37 °C for 5 minutes. Immunoblots were performed for ( a ) phosphorylated Src kinase family (Tyr416) and total Hck, Fgr, Lyn, phosphorylated and total p38, and β-Actin. Densitometry was performed for p-p38 and p-Src. ( b ) HCK, Fgr and Lyn were immunoprecipitated and tyrosine kinases activity assays and immunoblots were performed with the IP product. Data is represented as relative activity, corrected to absorbance for the wild type non-treated neutrophil group. N = 6 animal per group. Each measurement is the mean ± SEM. *Denotes a p value
    Figure Legend Snippet: Neutrophils from Pltp deficient mice has elevated tyrosine kinase activity of Hck. Neutrophils were collected from C57BL/6 and Pltp −/− mice. Neutrophils were exposed to AAT prior to fMLP at 37 °C for 5 minutes. Immunoblots were performed for ( a ) phosphorylated Src kinase family (Tyr416) and total Hck, Fgr, Lyn, phosphorylated and total p38, and β-Actin. Densitometry was performed for p-p38 and p-Src. ( b ) HCK, Fgr and Lyn were immunoprecipitated and tyrosine kinases activity assays and immunoblots were performed with the IP product. Data is represented as relative activity, corrected to absorbance for the wild type non-treated neutrophil group. N = 6 animal per group. Each measurement is the mean ± SEM. *Denotes a p value

    Techniques Used: Mouse Assay, Activity Assay, Western Blot, Immunoprecipitation

    Chemical inhibition of Src tyrosine kinase activity subdues neutrophil degranulation in Pltp −/− mice. Neutrophils were collected from C57BL/6 and Pltp −/− mice and were exposed to Src inhibitors, saracatinib or dasatinib, prior to fMLP at 37 °C for ( a ) 5 minutes or (b-cu unit3d) 0 minutes. ( b ) Immunoblots were performed for phosphorylated Src kinase family (Tyr416) and total Hck, Fgr, Lyn, phosphorylated and total p38, and β-Actin. Densitometry was performed for p-p38 and p-Src. ( c ) A reduced cytochrome c assay was used to determine production of O 2 − by neutrophils. ( c ) Cell-free supernatants were collected and primary (NE by ELISA and cathepsin G by substrate activity) granules were evaluated. N = 5 animal per group. Each measurement is the mean ± SEM. *Denotes a p value
    Figure Legend Snippet: Chemical inhibition of Src tyrosine kinase activity subdues neutrophil degranulation in Pltp −/− mice. Neutrophils were collected from C57BL/6 and Pltp −/− mice and were exposed to Src inhibitors, saracatinib or dasatinib, prior to fMLP at 37 °C for ( a ) 5 minutes or (b-cu unit3d) 0 minutes. ( b ) Immunoblots were performed for phosphorylated Src kinase family (Tyr416) and total Hck, Fgr, Lyn, phosphorylated and total p38, and β-Actin. Densitometry was performed for p-p38 and p-Src. ( c ) A reduced cytochrome c assay was used to determine production of O 2 − by neutrophils. ( c ) Cell-free supernatants were collected and primary (NE by ELISA and cathepsin G by substrate activity) granules were evaluated. N = 5 animal per group. Each measurement is the mean ± SEM. *Denotes a p value

    Techniques Used: Inhibition, Activity Assay, Mouse Assay, Western Blot, Enzyme-linked Immunosorbent Assay

    Proposed inhibition of neutrophil degranulation by AAT and PLTP. ( a ) Left panel, functional AAT (PiMM) prevents the cleavage of PLTP by proteases. Both AAT and PLTP prevent Src kinase activation to reduce superoxide production and degranulation. Right panel, AAT deficiency (PiZZ) results in reduced AAT circulation and elevated PLTP cleavage in the lungs. Reduced AAT and cleaved PLTP are less effective at minimizing superoxide production and degranulation in neutrophils. ( b ) Stimulation with fMLP or LTB 4 induces a signaling cascade leading to Hck activation and granule release. PLTP expression, AAT stimulation or Src inhibitors reduces Hck activation and degranulation. Fgr is known to regulate p38 activation but Hck could also contribute. Release of granulates further enhance PLTP cleavage and AAT complexes to serine proteases, which could further enhance degranulation.
    Figure Legend Snippet: Proposed inhibition of neutrophil degranulation by AAT and PLTP. ( a ) Left panel, functional AAT (PiMM) prevents the cleavage of PLTP by proteases. Both AAT and PLTP prevent Src kinase activation to reduce superoxide production and degranulation. Right panel, AAT deficiency (PiZZ) results in reduced AAT circulation and elevated PLTP cleavage in the lungs. Reduced AAT and cleaved PLTP are less effective at minimizing superoxide production and degranulation in neutrophils. ( b ) Stimulation with fMLP or LTB 4 induces a signaling cascade leading to Hck activation and granule release. PLTP expression, AAT stimulation or Src inhibitors reduces Hck activation and degranulation. Fgr is known to regulate p38 activation but Hck could also contribute. Release of granulates further enhance PLTP cleavage and AAT complexes to serine proteases, which could further enhance degranulation.

    Techniques Used: Inhibition, Functional Assay, Activation Assay, Expressing

    Related Articles

    Immunoprecipitation:

    Article Title: Nitroalkenes Suppress Lipopolysaccharide-Induced Signal Transducer and Activator of Transcription Signaling in Macrophages: A Critical Role of Mitogen-Activated Protein Kinase Phosphatase 1
    Article Snippet: .. The cell lysates were subjected to immunoprecipitation and immunoblot as previously described ( ) using antibodies of phospho-STAT1 (pSTAT1) (Tyr-701), pSTAT3 (Tyr-705), STAT1, and STAT3 (Cell Signaling Technology, Inc., Danvers, MA), MKP-1 and GAPDH (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), and iNOS, (BD Biosciences, San Jose, CA). .. For quantitation of the changes in STAT1 phosphorylation, the intensities of the bands representing STAT1 and pSTAT1 (Tyr-701) were measured by densitometry using an image scanner (EPSON GT-8000) and National Institutes of Health Image software.

    Article Title: Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages
    Article Snippet: .. After formaldehyde fixation, nuclei were isolated from macrophages, sheared, and immunoprecipitated using PU.1, IRF8, STAT1, or IRF1 antibodies. .. C1qB promoter sequences were sought in the precipitated chromatin fragments by PCR using primers spanning −167 and −31 bp, which contained the 53-bp element.

    Isolation:

    Article Title: Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages
    Article Snippet: .. After formaldehyde fixation, nuclei were isolated from macrophages, sheared, and immunoprecipitated using PU.1, IRF8, STAT1, or IRF1 antibodies. .. C1qB promoter sequences were sought in the precipitated chromatin fragments by PCR using primers spanning −167 and −31 bp, which contained the 53-bp element.

    Blocking Assay:

    Article Title: Colorectal cancer-derived microvesicles modulate differentiation of human monocytes to macrophages
    Article Snippet: .. Then, after blocking for 1 h at room temperature in Tris buffered saline (TBS) with 0.1 % Tween-20 (Sigma, St. Louis, MO) and 1 % bovine serum albumin (BSA, Sigma) the membranes were incubated overnight at 4 °C with rabbit polyclonal antibodies: anti-phopspho-STAT1 (Tyr701), anti-phospho-STAT3 (Tyr705), anti-phospho-STAT5 (Tyr694), anti-total STAT1, anti-total STAT3 and anti-total STAT5 (all antibodies were purchased from Cell Signaling, Danvers, MA) diluted 1:1000. .. After incubation, membranes were washed in TBS supplemented with BSA and Tween-20 and incubated for 1 h at room temperature with secondary goat anti-rabbit antibody (dilution 1:2500) conjugated with horseradish peroxidase (Cell Signaling).

    FLAG-tag:

    Article Title: Hemagglutinin of Influenza A Virus Antagonizes Type I Interferon (IFN) Responses by Inducing Degradation of Type I IFN Receptor 1
    Article Snippet: .. Antibodies against influenza viral NP, M2, IFNAR1 , and pIFNAR1 (S355/S539) were purchased from Abcam; antibody against influenza viral NS1 was purchased from Santa Cruz; antibodies against human GAPDH (glyceraldehyde-3-phosphate dehydrogenase), α subunit of eukaryotic initiation factor 2 (eIF2α), pelF2α, ISG15, ISG56, OAS1, TYK2, extracellular signal-regulated kinase (ERK), pERK, p38, p-p38, STAT1, pSTAT1, STAT2, pSTAT2, FLAG-tag, Myc-tag, and HA-tag were purchased from Cell Signaling Technology; antibodies against HA of IAV H1N1, HA of IAV H3N2, NA of IAV H5N1, and HA of avian IAV H5N1 were purchased from GeneTex. .. Mammalian expression plasmids encoding C-terminal 3×FLAG-tagged wild-type (WT) IFNAR1 and its phosphorylation-incompetent mutant (IFNAR1-S535/539A) were previously used ( , , ).

    Incubation:

    Article Title: Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages
    Article Snippet: .. After addition of sonicated salmon sperm DNA (25 μg), the preabsorbed lysate was then incubated overnight at 4 °C with resins loaded with the PU.1, IRF8, STAT1, or IRF1 antibodies or, as controls, resins loaded with the anti-His antibody or resins without antibody. ..

    Article Title: IL-10 regulates adult neurogenesis by modulating ERK and STAT3 activity
    Article Snippet: .. Membranes were then incubated with the following antibodies: phosphorylated STAT1 and STAT3 (all from Cell Signaling Technology, 1:1000); DCX (Cell Signaling Technology 1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA 1:500), Nestin (1:500, either from Merck-Millipore or BD Bioscience San Jose, CA, USA), Musashi (Millipore, 1:500), Mash1 (BD Bioscience, 1:500), total STAT-1 (BD transduction, 1:1000), total STAT-3 (BD Bioscience, 1:1000), Notch-ICD (Abcam, Cambridge, UK; 1:500), Numb (Abcam, 1:500); and Tubulin (Sigma, 1:50000), or Actin (Sigma, 1:50000) as loading controls. .. The list and dilution of primary antibodies was: pSTAT3ser727 (1:500), pERK1/2 (1:500), BIII-Tubulin (1:1000), DCX (1:1000), Nestin (1:200), Ki67 (Leica Microsystems, Wetzlar, Germany 1:1000), BrdU (AbCam, 1:400), Olig2 (Millipore, 1:400), GFAP (DAKO, 1:2000).

    Article Title: Colorectal cancer-derived microvesicles modulate differentiation of human monocytes to macrophages
    Article Snippet: .. Then, after blocking for 1 h at room temperature in Tris buffered saline (TBS) with 0.1 % Tween-20 (Sigma, St. Louis, MO) and 1 % bovine serum albumin (BSA, Sigma) the membranes were incubated overnight at 4 °C with rabbit polyclonal antibodies: anti-phopspho-STAT1 (Tyr701), anti-phospho-STAT3 (Tyr705), anti-phospho-STAT5 (Tyr694), anti-total STAT1, anti-total STAT3 and anti-total STAT5 (all antibodies were purchased from Cell Signaling, Danvers, MA) diluted 1:1000. .. After incubation, membranes were washed in TBS supplemented with BSA and Tween-20 and incubated for 1 h at room temperature with secondary goat anti-rabbit antibody (dilution 1:2500) conjugated with horseradish peroxidase (Cell Signaling).

    other:

    Article Title: Characterization and allergic role of IL-33-induced neutrophil polarization
    Article Snippet: The primary antibodies against p-JNK (Thr183/Tyr185), JNK, p-p38 (Thr180/Tyr182), p38, p-STAT1 (Ser727), STAT1, p-ERK (Thr202/Tyr204), ERK, p-p65 (Ser536), p65 and H3 were purchased from Cell Signaling Technology.

    Sonication:

    Article Title: Molecular Mechanisms for Synchronized Transcription of Three Complement C1q Subunit Genes in Dendritic Cells and Macrophages
    Article Snippet: .. After addition of sonicated salmon sperm DNA (25 μg), the preabsorbed lysate was then incubated overnight at 4 °C with resins loaded with the PU.1, IRF8, STAT1, or IRF1 antibodies or, as controls, resins loaded with the anti-His antibody or resins without antibody. ..

    Recombinant:

    Article Title: VSL#3 Probiotic Stimulates T-Cell Protein Tyrosine Phosphatase-Mediated Recovery of IFN-γ Induced Intestinal Epithelial Barrier Defects
    Article Snippet: .. VSL#3 packets with 450 billion bacteria per sachet (gift from Professor Claudio De Simone), human recombinant IFN-γ (Roche, Mannheim, Germany), cycloheximide (Sigma-Aldrich, St. Louis, MO, USA), monoclonal mouse anti-TCPTP antibody CF-4, which detects the 45-kilodalton and the 48-kilodalton isoforms (Calbiochem, San Diego, CA), mouse anti-TCPTP (Ab-1) antibody (EMD Millipore, Billerica, MA), anti-phospho-STAT1 (Tyr701 ), anti-STAT1, (Cell Signaling Technologies, Danvers, MA), Claudin-2, Occludin and ZO-1 (Invitrogen, Waltham, Massachusetts, USA) and monoclonal mouse anti-β-Actin (Sigma) were obtained from the sources noted. .. Millicell culture plate inserts were purchased from Millipore Corporation (Millipore, Bedford, MA).

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  • 94
    Cell Signaling Technology Inc p38
    Dose‐dependent reduction of age‐related cardiac inflammation by phenolic compounds. (a) Representative microphotographs of left ventricular sections of the heart stained with hematoxylin‐eosin showing young, SHAM, DMSO, PC 2.5, PC 5, PC 10, and PC 20 groups. (b) Histograms showing cardiomyocytes width (μm) and semiquantitative scores of inflammation ( n = 6–8 per group). (c) CRP and IL‐6 plasma concentrations in all groups ( n = 6–8 per group). (d) Representative western blot analyses for <t>p38</t> in rat cardiac tissue, normalized to GAPDH (in arbitrary units, a.u.) ( n = 3 for each protein and condition). * p
    P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1415 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p38/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc native p38 map kinase
    The Fyn-PKCδ signaling axis mediates <t>MAP</t> kinase activation in microglial cells. A , B , Immunoblot analysis demonstrated diminished LPS-induced <t>p38</t> and p44/42 (ERK) phosphorylation in Fyn −/− and PKCδ −/− microglia. * p
    Native P38 Map Kinase, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc sapk2a p38 α
    Fig. 3. Identification of the residues on TAB1 phosphorylated by <t>SAPK2a/p38α.</t> His-TAB1 was phosphorylated with active GST–SAPK2a/p38α and subjected to SDS–PAGE. The band corresponding to 32 P-labelled TAB1 was visualized by staining with Coomassie blue, excised and digested with trypsin. ( A ) The tryptic phosphopeptides were separated by HPLC on a Vydac C18 column equilibrated in 0.1% (v/v) trifluoroacetic acid. The column was developed with an acetonitrile gradient in 0.1% (v/v) trifluoroacetic acid (broken line). Radioactivity is indicated by the full line. ( B ) All the major 32 P-labelled peptides from (A) were pooled, further digested with the protease Asp-N and rechromatographed on the Vydac C18 column as in (A). Peptides D2, D3, D4 and D1 were then subjected to solid phase sequencing in ( C ), ( D ), ( E ) and ( F ).
    Sapk2a P38 α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Dose‐dependent reduction of age‐related cardiac inflammation by phenolic compounds. (a) Representative microphotographs of left ventricular sections of the heart stained with hematoxylin‐eosin showing young, SHAM, DMSO, PC 2.5, PC 5, PC 10, and PC 20 groups. (b) Histograms showing cardiomyocytes width (μm) and semiquantitative scores of inflammation ( n = 6–8 per group). (c) CRP and IL‐6 plasma concentrations in all groups ( n = 6–8 per group). (d) Representative western blot analyses for p38 in rat cardiac tissue, normalized to GAPDH (in arbitrary units, a.u.) ( n = 3 for each protein and condition). * p

    Journal: Aging Cell

    Article Title: Long‐term intake of phenolic compounds attenuates age‐related cardiac remodeling, et al. Long‐term intake of phenolic compounds attenuates age‐related cardiac remodeling

    doi: 10.1111/acel.12894

    Figure Lengend Snippet: Dose‐dependent reduction of age‐related cardiac inflammation by phenolic compounds. (a) Representative microphotographs of left ventricular sections of the heart stained with hematoxylin‐eosin showing young, SHAM, DMSO, PC 2.5, PC 5, PC 10, and PC 20 groups. (b) Histograms showing cardiomyocytes width (μm) and semiquantitative scores of inflammation ( n = 6–8 per group). (c) CRP and IL‐6 plasma concentrations in all groups ( n = 6–8 per group). (d) Representative western blot analyses for p38 in rat cardiac tissue, normalized to GAPDH (in arbitrary units, a.u.) ( n = 3 for each protein and condition). * p

    Article Snippet: The blots were then incubated overnight with gentle agitation at 4°C with primary antibodies: NFATc3 (Santa‐Cruz Biotechnology, Dallas, TX, USA; 1/500), phospho‐Ser165 NFATc3 (p‐NFATc3) (Abcam; 1/500), PP2B‐Aβ (C‐20) (Santa‐Cruz Biotechnology; 1/1,000), SOD1 (Abcam; 1/2000), SOD2 (Abcam; 1/4,000), Smad2 (Abcam; 1/500), phospho‐S467 Smad2 (p‐Smad2) (Abcam; 1/500), Smad3 (Abcam; 1/1,000), phospho‐S423+S425 Smad3 (p‐Smad3) (Abcam; 1/1,000), pan‐CAMKII (Cell Signaling Technology, MA, USA; 1/1,000), pan‐phospho CAMKII (Cell Signaling Technology; 1/1,000), troponin I (C‐4) (Santa‐Cruz Biotechnology; 1/500), GSK 3β (Cell Signaling Technology; 1/1,000), phospho‐GSK 3β (Cell Signaling Technology; 1/1,000), ERK1/2 (Abcam; 1/1,000), phospho‐ERK 1/2 (Abcam; 1/10,000), P38 (Cell Signaling Technology; 1/1,000), phospho‐Thr180/Tyr182 P38 (Cell Signaling Technology; 1/1,000), NF‐ƙB p65 (Ser536) (Cell Signaling Technology; 1/1,000), phospho‐NF‐ƙB p65 (Ser536) (93H1) (Cell Signaling Technology; 1/1,000), and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) (Abcam; 1/2,500 for all).

    Techniques: Staining, Western Blot

    Immunoactivity of phosphorylated p38 in IgA nephropathy patients according to the grade of interstitial fibrosis (original magnification x100). Control panel shows the phosphorylated p38 kidney tissue activity among patients who have no pathologic changes in kidney biopsy specimens. (Fig 1-A) The immunoactivity of phosphorylated p38 in tissues from IgA nephropathy patients with moderate interstitial fibrosis (Fig 1-D) was higher than that in tissues from patients with no interstitial fibrosis (Fig 1-B) or mild interstitial fibrosis (Fig 1-C). The immunoactivity of phosphorylated p38 in patients with a high degree of interstitial fibrosis was higher than immunoactivity in those with a low degree of kidney interstitial fibrosis (Fig 1-E; overall P

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Immunoactivity of phosphorylated p38 in IgA nephropathy patients according to the grade of interstitial fibrosis (original magnification x100). Control panel shows the phosphorylated p38 kidney tissue activity among patients who have no pathologic changes in kidney biopsy specimens. (Fig 1-A) The immunoactivity of phosphorylated p38 in tissues from IgA nephropathy patients with moderate interstitial fibrosis (Fig 1-D) was higher than that in tissues from patients with no interstitial fibrosis (Fig 1-B) or mild interstitial fibrosis (Fig 1-C). The immunoactivity of phosphorylated p38 in patients with a high degree of interstitial fibrosis was higher than immunoactivity in those with a low degree of kidney interstitial fibrosis (Fig 1-E; overall P

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques: Activity Assay

    Cellular mRNA expression of fibrosis-related markers after exposure to TGF-β in the presence of p38 MAPK inhibitor. (A) COL1 expression was increased after exposure to TGF-β (2 ng/ml). Treatment with the p38 MAPK inhibitor significantly reduced the cellular expression of COL1. (B) The decrease in the expression of fibronectin was not significant after concomitant exposure to TGF-β and p38 MAPK inhibitor. (C) Expression of periostin was increased after exposure to TGF-β (2 ng/ml). Treatment with p38 MAPK inhibitor reduced TGF-β-induced periostin expression. (n = 6 per group for each experiment, and in vitro experiments were repeated 3 times to confirm reproducibility) *, P

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Cellular mRNA expression of fibrosis-related markers after exposure to TGF-β in the presence of p38 MAPK inhibitor. (A) COL1 expression was increased after exposure to TGF-β (2 ng/ml). Treatment with the p38 MAPK inhibitor significantly reduced the cellular expression of COL1. (B) The decrease in the expression of fibronectin was not significant after concomitant exposure to TGF-β and p38 MAPK inhibitor. (C) Expression of periostin was increased after exposure to TGF-β (2 ng/ml). Treatment with p38 MAPK inhibitor reduced TGF-β-induced periostin expression. (n = 6 per group for each experiment, and in vitro experiments were repeated 3 times to confirm reproducibility) *, P

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques: Expressing, In Vitro

    Immunohistochemical staining showing the expression of phosphorylated p38 in the mouse renal tubulointerstitial area after UUO and administration of p38 MAPK inhibitor. Scale bar = 50 μm (original magnification x300). (A) Sham-operated group. (B) UUO-operated group administered 20% DMSO vehicle. (C) UUO-operated group administered 5 mg/kg/day p38 MAPK inhibitor. (D) UUO-operated group administered 10 mg/kg/day p38 MAPK inhibitor. Tubulointerstitial expression of phosphorylated p38 decreased significantly in the groups treated with p38 MAPK inhibitor. UUO, unilateral ureteral obstruction.

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Immunohistochemical staining showing the expression of phosphorylated p38 in the mouse renal tubulointerstitial area after UUO and administration of p38 MAPK inhibitor. Scale bar = 50 μm (original magnification x300). (A) Sham-operated group. (B) UUO-operated group administered 20% DMSO vehicle. (C) UUO-operated group administered 5 mg/kg/day p38 MAPK inhibitor. (D) UUO-operated group administered 10 mg/kg/day p38 MAPK inhibitor. Tubulointerstitial expression of phosphorylated p38 decreased significantly in the groups treated with p38 MAPK inhibitor. UUO, unilateral ureteral obstruction.

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques: Immunohistochemistry, Staining, Expressing

    Microscopic morphological changes in hTECs after treatment with p38 MAPK inhibitor. (A) Control hTECs show a normal ellipsoid shape and clear contour. (B) Exposure to TGF-β (2 ng/ml) induced bursting and speckled morphological changes in hTECs. (C) Exposure to TGF-β (2 ng/ml) in the presence of p38 MAPK inhibitor (2 μM). (D) Exposure to TGF-β (2 ng/ml) in the presence of p38 MAPK inhibitor (4 μM).

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Microscopic morphological changes in hTECs after treatment with p38 MAPK inhibitor. (A) Control hTECs show a normal ellipsoid shape and clear contour. (B) Exposure to TGF-β (2 ng/ml) induced bursting and speckled morphological changes in hTECs. (C) Exposure to TGF-β (2 ng/ml) in the presence of p38 MAPK inhibitor (2 μM). (D) Exposure to TGF-β (2 ng/ml) in the presence of p38 MAPK inhibitor (4 μM).

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques:

    Fibrosis-related renal protein expression as assessed with Western blot. (A) Expression of αSMA did not decrease after p38 MAPK inhibitor administration. (B) Expression of COL1 decreased significantly after administration of 10 mg/kg/day p38 MAPK inhibitor. (C) Expression of phosphorylated p38 MAPK decreased significantly after administration of 10 mg/kg/day p38 MAPK inhibitor. αSMA, α-smooth muscle actin; COL1, type 1 collagen. † , P

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Fibrosis-related renal protein expression as assessed with Western blot. (A) Expression of αSMA did not decrease after p38 MAPK inhibitor administration. (B) Expression of COL1 decreased significantly after administration of 10 mg/kg/day p38 MAPK inhibitor. (C) Expression of phosphorylated p38 MAPK decreased significantly after administration of 10 mg/kg/day p38 MAPK inhibitor. αSMA, α-smooth muscle actin; COL1, type 1 collagen. † , P

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques: Expressing, Western Blot

    Histologic Masson's trichrome stain findings of renal fibrosis after UUO and administration of a p38 MAPK inhibitor. Scale bar = 100 μm (original magnification x100). Mice from the sham-operated control group (A) show sparse tubulointerstitial fibrosis. UUO-operated group with administered 20% DMSO vehicle (B) shows prominent tubulointerstitial fibrosis. UUO-operated group administered p38 MAPK inhibitor at 5 mg/kg/day (C). UUO-operated group administered p38 MAPK inhibitor at 10 mg/kg/day (D). The extent of tubulointerstitial fibrosis in the groups administered p38 MAPK inhibitor was lower than that in the vehicle group (E). *, P

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Histologic Masson's trichrome stain findings of renal fibrosis after UUO and administration of a p38 MAPK inhibitor. Scale bar = 100 μm (original magnification x100). Mice from the sham-operated control group (A) show sparse tubulointerstitial fibrosis. UUO-operated group with administered 20% DMSO vehicle (B) shows prominent tubulointerstitial fibrosis. UUO-operated group administered p38 MAPK inhibitor at 5 mg/kg/day (C). UUO-operated group administered p38 MAPK inhibitor at 10 mg/kg/day (D). The extent of tubulointerstitial fibrosis in the groups administered p38 MAPK inhibitor was lower than that in the vehicle group (E). *, P

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques: Staining, Mouse Assay

    Fibrosis-related mRNA expression as assessed with RT-PCR. (A) Expression of αSMA was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (B) Expression of COL1 was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (C) Expression of fibronectin was decreased in the 5 and 10 mg/kg/day p38 MAPK inhibitor groups, with a dose-dependent pattern. (D) Expression of p38 MAPK was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (E) Expression of MCP-1 was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (F) The decrease in periostin expression was not statistically significant. αSMA, α-smooth muscle actin; COL1, type 1 collagen; MCP-1, monocyte chemoattractant protein-1; UUO, unilateral ureteral obstruction. *, P

    Journal: PLoS ONE

    Article Title: p38 MAPK activity is associated with the histological degree of interstitial fibrosis in IgA nephropathy patients

    doi: 10.1371/journal.pone.0213981

    Figure Lengend Snippet: Fibrosis-related mRNA expression as assessed with RT-PCR. (A) Expression of αSMA was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (B) Expression of COL1 was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (C) Expression of fibronectin was decreased in the 5 and 10 mg/kg/day p38 MAPK inhibitor groups, with a dose-dependent pattern. (D) Expression of p38 MAPK was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (E) Expression of MCP-1 was decreased in the 10 mg/kg/day p38 MAPK inhibitor group. (F) The decrease in periostin expression was not statistically significant. αSMA, α-smooth muscle actin; COL1, type 1 collagen; MCP-1, monocyte chemoattractant protein-1; UUO, unilateral ureteral obstruction. *, P

    Article Snippet: In the groups with treated with 5 mg/kg/day and 10 mg/kg/day p38 MAPK inhibitor, the level of phosphorylated p38 showed a tendency to decrease compared with that in the vehicle group ( , P = 0.20 vehicle group vs. p38 MAPK inhibitor 5 mg/kg/day group).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    The Fyn-PKCδ signaling axis mediates MAP kinase activation in microglial cells. A , B , Immunoblot analysis demonstrated diminished LPS-induced p38 and p44/42 (ERK) phosphorylation in Fyn −/− and PKCδ −/− microglia. * p

    Journal: The Journal of Neuroscience

    Article Title: Fyn Kinase Regulates Microglial Neuroinflammatory Responses in Cell Culture and Animal Models of Parkinson's Disease

    doi: 10.1523/JNEUROSCI.0302-15.2015

    Figure Lengend Snippet: The Fyn-PKCδ signaling axis mediates MAP kinase activation in microglial cells. A , B , Immunoblot analysis demonstrated diminished LPS-induced p38 and p44/42 (ERK) phosphorylation in Fyn −/− and PKCδ −/− microglia. * p

    Article Snippet: Antibodies against rabbit p-Src family kinase Y416 (p-Y416 SFK), native p65, p-p38 MAP kinase, native p38 MAP kinase, p-p44/42 MAP kinase (p-ERK), and native p44/42 MAP kinase (ERK) were purchased from Cell Signaling Technology.

    Techniques: Activation Assay

    Fig. 3. Identification of the residues on TAB1 phosphorylated by SAPK2a/p38α. His-TAB1 was phosphorylated with active GST–SAPK2a/p38α and subjected to SDS–PAGE. The band corresponding to 32 P-labelled TAB1 was visualized by staining with Coomassie blue, excised and digested with trypsin. ( A ) The tryptic phosphopeptides were separated by HPLC on a Vydac C18 column equilibrated in 0.1% (v/v) trifluoroacetic acid. The column was developed with an acetonitrile gradient in 0.1% (v/v) trifluoroacetic acid (broken line). Radioactivity is indicated by the full line. ( B ) All the major 32 P-labelled peptides from (A) were pooled, further digested with the protease Asp-N and rechromatographed on the Vydac C18 column as in (A). Peptides D2, D3, D4 and D1 were then subjected to solid phase sequencing in ( C ), ( D ), ( E ) and ( F ).

    Journal: The EMBO Journal

    Article Title: Feedback control of the protein kinase TAK1 by SAPK2a/p38?

    doi: 10.1093/emboj/cdg552

    Figure Lengend Snippet: Fig. 3. Identification of the residues on TAB1 phosphorylated by SAPK2a/p38α. His-TAB1 was phosphorylated with active GST–SAPK2a/p38α and subjected to SDS–PAGE. The band corresponding to 32 P-labelled TAB1 was visualized by staining with Coomassie blue, excised and digested with trypsin. ( A ) The tryptic phosphopeptides were separated by HPLC on a Vydac C18 column equilibrated in 0.1% (v/v) trifluoroacetic acid. The column was developed with an acetonitrile gradient in 0.1% (v/v) trifluoroacetic acid (broken line). Radioactivity is indicated by the full line. ( B ) All the major 32 P-labelled peptides from (A) were pooled, further digested with the protease Asp-N and rechromatographed on the Vydac C18 column as in (A). Peptides D2, D3, D4 and D1 were then subjected to solid phase sequencing in ( C ), ( D ), ( E ) and ( F ).

    Article Snippet: Antibodies that recognize TAB2 were obtained from Santa Cruz (Santa Cruz, CA) and antibodies that recognize all forms of SAPK2a/p38α or IκB, an antibody that recognizes IκB phosphorylated at Ser32 and an antibody that recognizes the active phosphorylated forms of ERK1 and ERK2 were purchased from Cell Signalling (Beverly, MA).

    Techniques: SDS Page, Staining, High Performance Liquid Chromatography, Radioactivity, Sequencing

    Fig. 11. Schematic representation of the feedback control of TAK1 activity by SAPK2a/p38α and its implication for the regulation of JNK and NF-κB. ( A ) SAPK2a/p38α downregulates TAK1, probably via the phosphorylation of TAB1. ( B ) The inhibition of SAPK2a/p38α by SB 203580 (or other inhibitors of this protein kinase) abolishes this feedback control of TAK1, causing upregulation of the JNK and IKK pathways (illustrated by the thicker arrows).

    Journal: The EMBO Journal

    Article Title: Feedback control of the protein kinase TAK1 by SAPK2a/p38?

    doi: 10.1093/emboj/cdg552

    Figure Lengend Snippet: Fig. 11. Schematic representation of the feedback control of TAK1 activity by SAPK2a/p38α and its implication for the regulation of JNK and NF-κB. ( A ) SAPK2a/p38α downregulates TAK1, probably via the phosphorylation of TAB1. ( B ) The inhibition of SAPK2a/p38α by SB 203580 (or other inhibitors of this protein kinase) abolishes this feedback control of TAK1, causing upregulation of the JNK and IKK pathways (illustrated by the thicker arrows).

    Article Snippet: Antibodies that recognize TAB2 were obtained from Santa Cruz (Santa Cruz, CA) and antibodies that recognize all forms of SAPK2a/p38α or IκB, an antibody that recognizes IκB phosphorylated at Ser32 and an antibody that recognizes the active phosphorylated forms of ERK1 and ERK2 were purchased from Cell Signalling (Beverly, MA).

    Techniques: Activity Assay, Inhibition

    Fig. 9. The TNF-α-induced activation of TAK1 is enhanced in embryonic mouse fibroblasts deficient in SAPK2a/p38α. Immortalized embryonic fibroblasts from wild-type (WT, open bars) and SAPK2a/p38α knockout (KO, black bars) mice were treated for 1 h with or without 10 µM SB 203580, then stimulated for 10 min with 10 ng/ml of mouse TNF-α, for 20 min with 20 ng/ml of mouse IL-1 or for 30 min with 0.5 M sorbitol, and the cells lysed. ( A ) The complexes containing TAK1 were immunoprecipitated from the cell lysates (0.15 mg of protein) using a TAB1-specific antibody, and the activity of TAK1 in the cell lysates determined (see Materials and methods). The results are expressed as the mean ± SEM for three separate experiments with all assays performed in duplicate. Results are shown as a percentage of the activity of TAK1 in stimulated wild-type cells measured without pre-incubation with SB 203580. ( B ) The TAK1 complexes immunoprecipitated from the lysates of cells stimulated with TNF-α were prepared as in (A), then subjected to SDS–PAGE followed by immunoblotting with antibodies that recognize TAK1, TAB1 and TAB2. Cell lysate (20 µg of protein) was also subjected to SDS–PAGE followed by immunoblotting for SAPK2a/p38α ( C ). TAK1 complexes immunoprecipitated from the lysates of cells stimulated with TNF-α or IL-1 were prepared as in (A), and then analysed as in (B), except that the gels were immunoblotted with antibodies that recognize TAB1 phosphorylated at Ser423, Thr431 or Ser438.

    Journal: The EMBO Journal

    Article Title: Feedback control of the protein kinase TAK1 by SAPK2a/p38?

    doi: 10.1093/emboj/cdg552

    Figure Lengend Snippet: Fig. 9. The TNF-α-induced activation of TAK1 is enhanced in embryonic mouse fibroblasts deficient in SAPK2a/p38α. Immortalized embryonic fibroblasts from wild-type (WT, open bars) and SAPK2a/p38α knockout (KO, black bars) mice were treated for 1 h with or without 10 µM SB 203580, then stimulated for 10 min with 10 ng/ml of mouse TNF-α, for 20 min with 20 ng/ml of mouse IL-1 or for 30 min with 0.5 M sorbitol, and the cells lysed. ( A ) The complexes containing TAK1 were immunoprecipitated from the cell lysates (0.15 mg of protein) using a TAB1-specific antibody, and the activity of TAK1 in the cell lysates determined (see Materials and methods). The results are expressed as the mean ± SEM for three separate experiments with all assays performed in duplicate. Results are shown as a percentage of the activity of TAK1 in stimulated wild-type cells measured without pre-incubation with SB 203580. ( B ) The TAK1 complexes immunoprecipitated from the lysates of cells stimulated with TNF-α were prepared as in (A), then subjected to SDS–PAGE followed by immunoblotting with antibodies that recognize TAK1, TAB1 and TAB2. Cell lysate (20 µg of protein) was also subjected to SDS–PAGE followed by immunoblotting for SAPK2a/p38α ( C ). TAK1 complexes immunoprecipitated from the lysates of cells stimulated with TNF-α or IL-1 were prepared as in (A), and then analysed as in (B), except that the gels were immunoblotted with antibodies that recognize TAB1 phosphorylated at Ser423, Thr431 or Ser438.

    Article Snippet: Antibodies that recognize TAB2 were obtained from Santa Cruz (Santa Cruz, CA) and antibodies that recognize all forms of SAPK2a/p38α or IκB, an antibody that recognizes IκB phosphorylated at Ser32 and an antibody that recognizes the active phosphorylated forms of ERK1 and ERK2 were purchased from Cell Signalling (Beverly, MA).

    Techniques: Activation Assay, Knock-Out, Mouse Assay, Immunoprecipitation, Activity Assay, Incubation, SDS Page

    Fig. 1. TAB1 interacts specifically with SAPK2a/p38α. HEK 293 cells were transfected with plasmids coding for GST–TAB1 and SAPK2a/p38α, SAPK2b/p38β2, SAPK3/p38γ, SAPK4/p38δ, MKK3 or MKK6 fused to an HA epitope tag. After 36 h, the cells were lysed and the cell lysate (20 µg of protein) denatured in SDS, subjected to SDS–PAGE, transferred onto nitrocellulose membrane and the membrane probed with an antibody specific for the HA epitope to detect the expression of each protein kinase or with an anti-GST antibody to detect the level of expression of GST–TAB1. The GST–TAB1 in the cell lysates was then affinity purified on glutathione–Sepharose and proteins binding to this support were subjected to SDS–PAGE and immunoblotting with the anti-HA antibody to investigate the binding of each protein kinase to TAB1.

    Journal: The EMBO Journal

    Article Title: Feedback control of the protein kinase TAK1 by SAPK2a/p38?

    doi: 10.1093/emboj/cdg552

    Figure Lengend Snippet: Fig. 1. TAB1 interacts specifically with SAPK2a/p38α. HEK 293 cells were transfected with plasmids coding for GST–TAB1 and SAPK2a/p38α, SAPK2b/p38β2, SAPK3/p38γ, SAPK4/p38δ, MKK3 or MKK6 fused to an HA epitope tag. After 36 h, the cells were lysed and the cell lysate (20 µg of protein) denatured in SDS, subjected to SDS–PAGE, transferred onto nitrocellulose membrane and the membrane probed with an antibody specific for the HA epitope to detect the expression of each protein kinase or with an anti-GST antibody to detect the level of expression of GST–TAB1. The GST–TAB1 in the cell lysates was then affinity purified on glutathione–Sepharose and proteins binding to this support were subjected to SDS–PAGE and immunoblotting with the anti-HA antibody to investigate the binding of each protein kinase to TAB1.

    Article Snippet: Antibodies that recognize TAB2 were obtained from Santa Cruz (Santa Cruz, CA) and antibodies that recognize all forms of SAPK2a/p38α or IκB, an antibody that recognizes IκB phosphorylated at Ser32 and an antibody that recognizes the active phosphorylated forms of ERK1 and ERK2 were purchased from Cell Signalling (Beverly, MA).

    Techniques: Transfection, SDS Page, Expressing, Affinity Purification, Binding Assay

    Fig. 2. TAB1 is phosphorylated stoichiometrically by SAPK2a/p38α. Bacterially expressed His-TAB1 was phosphorylated at 30°C with 2 U/ml GST–SAPK2a/p38α and Mg [γ- 32 P]ATP. At the times indicated, an aliquot of the reaction was denatured in SDS and subjected to SDS–PAGE. The gel was stained with Coomassie blue and destained ( B ). The band corresponding to His-TAB1 was excised and analysed by Cerenkov counting to quantitate the incorporation of phosphate into TAB1 ( A ).

    Journal: The EMBO Journal

    Article Title: Feedback control of the protein kinase TAK1 by SAPK2a/p38?

    doi: 10.1093/emboj/cdg552

    Figure Lengend Snippet: Fig. 2. TAB1 is phosphorylated stoichiometrically by SAPK2a/p38α. Bacterially expressed His-TAB1 was phosphorylated at 30°C with 2 U/ml GST–SAPK2a/p38α and Mg [γ- 32 P]ATP. At the times indicated, an aliquot of the reaction was denatured in SDS and subjected to SDS–PAGE. The gel was stained with Coomassie blue and destained ( B ). The band corresponding to His-TAB1 was excised and analysed by Cerenkov counting to quantitate the incorporation of phosphate into TAB1 ( A ).

    Article Snippet: Antibodies that recognize TAB2 were obtained from Santa Cruz (Santa Cruz, CA) and antibodies that recognize all forms of SAPK2a/p38α or IκB, an antibody that recognizes IκB phosphorylated at Ser32 and an antibody that recognizes the active phosphorylated forms of ERK1 and ERK2 were purchased from Cell Signalling (Beverly, MA).

    Techniques: SDS Page, Staining

    Fig. 4. Development of phospho-specific antibodies recognizing each of the three phosphorylation sites on TAB1. Bacterially expressed His-TAB1 (0.5 µg) was phosphorylated with SAPK2a/p38α (P) or left unphosphorylated (U). The proteins were then separated by SDS–PAGE, transferred to nitrocellulose and probed with each of the three phospho-specific antibodies in the absence (none) or presence of 10 µg/ml of one of the three phosphopeptide immunogens (termed competitor peptide). Each antibody also contained 10 µg/ml of the unphosphorylated form of the relevant peptide immunogen to neutralize any antibodies present that recognize the unphosphorylated species. The bottom panel shows the amount of His-TAB1 in each gel lane stained with Coomassie Blue.

    Journal: The EMBO Journal

    Article Title: Feedback control of the protein kinase TAK1 by SAPK2a/p38?

    doi: 10.1093/emboj/cdg552

    Figure Lengend Snippet: Fig. 4. Development of phospho-specific antibodies recognizing each of the three phosphorylation sites on TAB1. Bacterially expressed His-TAB1 (0.5 µg) was phosphorylated with SAPK2a/p38α (P) or left unphosphorylated (U). The proteins were then separated by SDS–PAGE, transferred to nitrocellulose and probed with each of the three phospho-specific antibodies in the absence (none) or presence of 10 µg/ml of one of the three phosphopeptide immunogens (termed competitor peptide). Each antibody also contained 10 µg/ml of the unphosphorylated form of the relevant peptide immunogen to neutralize any antibodies present that recognize the unphosphorylated species. The bottom panel shows the amount of His-TAB1 in each gel lane stained with Coomassie Blue.

    Article Snippet: Antibodies that recognize TAB2 were obtained from Santa Cruz (Santa Cruz, CA) and antibodies that recognize all forms of SAPK2a/p38α or IκB, an antibody that recognizes IκB phosphorylated at Ser32 and an antibody that recognizes the active phosphorylated forms of ERK1 and ERK2 were purchased from Cell Signalling (Beverly, MA).

    Techniques: SDS Page, Staining