anti-p38 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Cell Signaling Technology Inc p38 mapk
    Pannexin 1 (Panx1) blockade reduces <t>p38</t> mitogen-activated protein kinase <t>(MAPK)</t> activation in hippocampal slices of AD transgenic (Tg) mice. (A) Representative blots of p-p38MAPK and t-p38MAPK expression in whole hippocampal homogenates of wild-type (Wt) and Tg mice at 6 months old (m.o.) in the absence (Wt, black; Tg, red) or the presence of 100 μM probenecid (PBN; Wt+PBN, gray; Tg+PBN, blue; left panel). Western blot analysis of phosphorylated p38MAPK levels normalized to total p38MAPK levels (right panel). One-way ANOVA ( F (3,20) = 15.95; p
    P38 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 7364 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p38 mapk/product/Cell Signaling Technology Inc
    Average 99 stars, based on 7364 article reviews
    Price from $9.99 to $1999.99
    p38 mapk - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Cell Signaling Technology Inc anti phospho p38
    Nck1, but not Nck2, is essential for activation of Erk1/2 and MEK1/2 proteins. Cells from each monoclonal group were untreated or treated with soluble CD3 antibody (1 μg/ml) at various time points. Cell lysates were subjected to immunoblot with anti-phospho-Erk1/2 antibody (A) . Below, the quantified signal intensity of the pErk1/2 was normalized to its total kinase and this value was relative to that in the unstimulated control cells (Mock), set as 1 (black dashed line) and plotted in bar graph. B) Immunoblot analysis of phospho-MEK1/2 from the lysates of cells that were stimulated as in A . Below, signal intensity was quantified and presented as a ratio of p-MEK1/2 to actin relative to that in unstimulated control cells (Mock), set as 1 (black dashed line). C) Immunoblot analysis of <t>phospho-p38.</t> Below, signal intensity was quantified and presented as described in A . D) . Polyclonal cells from Mock, Nck1- and Nck2-knockdown cells were stimulated as in A . Below, signal intensity was quantified and presented as described in A . E) Both Nck1 and Nck2 genes were co-silenced in Jurkat T cells. Nck expression levels were analysed by immunoblotting using indicated antibodies. F) The expression levels of surface CD3 molecules on double knockdown Nck1/2 were measured by flow cytometry. G) Cells from each monoclonal group, Nck1-, Nck2- and Nck1/2-knockdown cells were treated as in A for 0, 3 and 30 min. Lysates were subjected to immunoblot with anti-phospho-Erk1/2 antibody. Below, signal intensity was quantified and presented as described in A . Data are representative of at least two independent experiments.
    Anti Phospho P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 3888 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho p38/product/Cell Signaling Technology Inc
    Average 92 stars, based on 3888 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc p p38 mapk
    The relative mRNA expressions of STE20, TRAF6, IGF1R, and <t>p38</t> <t>MAPK</t> were shown at the 3 th , 7 th , 14 th , and 21 th day. The data are expressed as mean ± SD. ∗P
    P P38 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1398 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p p38 mapk/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1398 article reviews
    Price from $9.99 to $1999.99
    p p38 mapk - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc p38
    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 <t>p38.</t> 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.
    P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 14610 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p38/product/Cell Signaling Technology Inc
    Average 99 stars, based on 14610 article reviews
    Price from $9.99 to $1999.99
    p38 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Cell Signaling Technology Inc anti phospho p38 mapk
    DHX9 is not required for TLR2-stimulated NF-κB or <t>p38</t> <t>MAPK</t> activation. THP-1 cells expressing doxycycline-inducible DHX9-specific shRNA (G2) or NSC shRNA, were incubated with 1 μg/ml of doxycycline for 72 h, and then stimulated with 2
    Anti Phospho P38 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1271 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho p38 mapk/product/Cell Signaling Technology Inc
    Average 93 stars, based on 1271 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 mapk - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology anti p38
    Activation of MAPKs on human breast carcinoma MCF-7 cells treated with PMA. Cells were incubated in the presence of PMA (50 ng/ml) during the indicated times, and the activation status of ERK1/2, <t>p38,</t> and JNK determined using phospho-specific anti-ERK1/2
    Anti P38, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p38/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1202 article reviews
    Price from $9.99 to $1999.99
    anti p38 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    91
    Cell Signaling Technology Inc rabbit anti phospho p38
    The <t>p38</t> cascade is up-regulated in patients with relapsing–remitting multiple sclerosis (RR-MS): (a) Graph represents the median fluorescence intensity (MFI) of p38 total protein (left) and phosphorylated protein (right) in peripheral blood mononuclear
    Rabbit Anti Phospho P38, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 544 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti phospho p38/product/Cell Signaling Technology Inc
    Average 91 stars, based on 544 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho p38 - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc anti p p38 thr180 tyr182
    The <t>p38</t> cascade is up-regulated in patients with relapsing–remitting multiple sclerosis (RR-MS): (a) Graph represents the median fluorescence intensity (MFI) of p38 total protein (left) and phosphorylated protein (right) in peripheral blood mononuclear
    Anti P P38 Thr180 Tyr182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p p38 thr180 tyr182/product/Cell Signaling Technology Inc
    Average 86 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    anti p p38 thr180 tyr182 - by Bioz Stars, 2020-09
    86/100 stars
      Buy from Supplier

    p38  (Abcam)
    97
    Abcam p38
    APIM-peptide increases the effects of docetaxel on apoptosis and cellular signaling ( A ) Cell cycle analysis and fraction of apoptotic (A) vs necrotic (N) cells (values from contour plots are given). ( B ) Relative expression of phosphorylated Akt, ERK1, ERK2, S6K, and <t>p38,</t> cleaved-PARP, and caspase 3. For all experiments, PC3 and Du145 cells were treated with APIM-peptide (14 μg/mL, green), docetaxel (2.5 ng/mL: PC3; 1.3 ng/mL: Du145; 5 ng/mL, blue) and the combination of these (black) for 24 hours prior to the analysis. The protein levels (B) were adjusted for loading differences (β-tubulin) and normalized against untreated cells, and additionally for total protein levels for the phosphorylated proteins. Data are from three biological replicas (different symbols represents extracts acquired on different days). Statistically significant differences ( * p
    P38, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 879 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p38/product/Abcam
    Average 97 stars, based on 879 article reviews
    Price from $9.99 to $1999.99
    p38 - by Bioz Stars, 2020-09
    97/100 stars
      Buy from Supplier

    95
    Abcam anti p38
    Activated <t>p38</t> downregulates the stem cell properties of NSCLC cells ( A ) Flow cytometry showing the percentage of the side population detected by Hoechst33342 staining in A549 and H460 cells transduced with vector control (BP), MKK3E or MKK6E and H460 and H1299 cells transduced with vector control (BH), MKK3A or MKK6A. Bar graphs show quantification of the percentage of side population. ( B ) Sphere formation assay of A549 and H460 cells transduced with vector control (BP), MKK3E or MKK6E and H460 and H1299 cells transduced with vector control (BH), MKK3A or MKK6A. Scar bar, 100 μm. Bar graphs show quantifications of the number and diameter of the spheres, respectively. * indicates P
    Anti P38, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 398 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p38/product/Abcam
    Average 95 stars, based on 398 article reviews
    Price from $9.99 to $1999.99
    anti p38 - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    Image Search Results


    Pannexin 1 (Panx1) blockade reduces p38 mitogen-activated protein kinase (MAPK) activation in hippocampal slices of AD transgenic (Tg) mice. (A) Representative blots of p-p38MAPK and t-p38MAPK expression in whole hippocampal homogenates of wild-type (Wt) and Tg mice at 6 months old (m.o.) in the absence (Wt, black; Tg, red) or the presence of 100 μM probenecid (PBN; Wt+PBN, gray; Tg+PBN, blue; left panel). Western blot analysis of phosphorylated p38MAPK levels normalized to total p38MAPK levels (right panel). One-way ANOVA ( F (3,20) = 15.95; p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Acute Pannexin 1 Blockade Mitigates Early Synaptic Plasticity Defects in a Mouse Model of Alzheimer’s Disease

    doi: 10.3389/fncel.2020.00046

    Figure Lengend Snippet: Pannexin 1 (Panx1) blockade reduces p38 mitogen-activated protein kinase (MAPK) activation in hippocampal slices of AD transgenic (Tg) mice. (A) Representative blots of p-p38MAPK and t-p38MAPK expression in whole hippocampal homogenates of wild-type (Wt) and Tg mice at 6 months old (m.o.) in the absence (Wt, black; Tg, red) or the presence of 100 μM probenecid (PBN; Wt+PBN, gray; Tg+PBN, blue; left panel). Western blot analysis of phosphorylated p38MAPK levels normalized to total p38MAPK levels (right panel). One-way ANOVA ( F (3,20) = 15.95; p

    Article Snippet: For both cases, 40 μg of protein per lane were resolved by 10% SDS-polyacrylamide gel electrophoresis (PAGE), followed by immunoblotting on polyvinylidene fluoride (PVDF) membranes (BioRad, Berkeley, CA, USA) and probed with specific antibodies against Panx1 (rabbit anti-Panx1, ABN242 Merck; 1:1,000), PSD-95 (mouse anti-PSD-95, MAB1596 Merck, 1:1,000), synaptophysin (goat anti-SYP, sc-9116, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:2,000), p38 MAPK (rabbit anti-p38MAPK, #9212, Cell Signaling Technology, Danvers, MA, USA; 1:1,000), phospho-p38 MAPK (rabbit anti-Thr180/Tyr182, #9211, Cell Signaling Technology, Danvers, MA, USA; 1:1,000), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse anti-GAPDH, sc-47724, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:1,000).

    Techniques: Activation Assay, Transgenic Assay, Mouse Assay, Expressing, Western Blot

    Proposed model for pannexin 1 (Panx1)’s role in AD synaptotoxicity. Amyloid β peptide (Aβ) is generated by the proteolytic cleavage of the amyloid precursor protein (APP). This processing is executed by the consecutive actions of β-secretase (not shown) and γ-secretase resulting in the release of a soluble β-cleaved APP fragment (sAPPβ) and the Aβ, respectively. Aβ can form soluble oligomeric aggregates (sAβos) that easily diffuse and bind to several postsynaptic partners including N-methyl-D-aspartate receptors (NMDARs) and type 1 metabotropic glutamate receptor 5 (mGluR5), enhancing glutamatergic transmission and promoting NMDAR/mGluR5-mediated activation of Panx1 channels. Panx1 overactivity favors signaling cascades that promote the activation (phosphorylation) of p38-MAPK (p38MAPK), a kinase involved in the early stages of the Aβ-induced synaptic dysfunction in AD. In turn, p38MAPK promotes Aβ production and accumulation, further increasing Panx1 overactivity and producing a “positive loop” that amplifies the Aβ-induced neurotoxicity. Congruently with this mechanism, the inhibition of Panx1 with probenecid (PBN) reverses p38MAPK activation and improves LTP/LTD and structural impairments in the AD model brain. Similarly, inhibition of p38MAPK with SB208035 (SB) or the inhibition of γ-secretase-with N-[N-(3,5-difluorophenacetyl-l-alanyl)]-S-phenylglycine t-butyl ester (DAPT) reduces Panx1 overactivity.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Acute Pannexin 1 Blockade Mitigates Early Synaptic Plasticity Defects in a Mouse Model of Alzheimer’s Disease

    doi: 10.3389/fncel.2020.00046

    Figure Lengend Snippet: Proposed model for pannexin 1 (Panx1)’s role in AD synaptotoxicity. Amyloid β peptide (Aβ) is generated by the proteolytic cleavage of the amyloid precursor protein (APP). This processing is executed by the consecutive actions of β-secretase (not shown) and γ-secretase resulting in the release of a soluble β-cleaved APP fragment (sAPPβ) and the Aβ, respectively. Aβ can form soluble oligomeric aggregates (sAβos) that easily diffuse and bind to several postsynaptic partners including N-methyl-D-aspartate receptors (NMDARs) and type 1 metabotropic glutamate receptor 5 (mGluR5), enhancing glutamatergic transmission and promoting NMDAR/mGluR5-mediated activation of Panx1 channels. Panx1 overactivity favors signaling cascades that promote the activation (phosphorylation) of p38-MAPK (p38MAPK), a kinase involved in the early stages of the Aβ-induced synaptic dysfunction in AD. In turn, p38MAPK promotes Aβ production and accumulation, further increasing Panx1 overactivity and producing a “positive loop” that amplifies the Aβ-induced neurotoxicity. Congruently with this mechanism, the inhibition of Panx1 with probenecid (PBN) reverses p38MAPK activation and improves LTP/LTD and structural impairments in the AD model brain. Similarly, inhibition of p38MAPK with SB208035 (SB) or the inhibition of γ-secretase-with N-[N-(3,5-difluorophenacetyl-l-alanyl)]-S-phenylglycine t-butyl ester (DAPT) reduces Panx1 overactivity.

    Article Snippet: For both cases, 40 μg of protein per lane were resolved by 10% SDS-polyacrylamide gel electrophoresis (PAGE), followed by immunoblotting on polyvinylidene fluoride (PVDF) membranes (BioRad, Berkeley, CA, USA) and probed with specific antibodies against Panx1 (rabbit anti-Panx1, ABN242 Merck; 1:1,000), PSD-95 (mouse anti-PSD-95, MAB1596 Merck, 1:1,000), synaptophysin (goat anti-SYP, sc-9116, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:2,000), p38 MAPK (rabbit anti-p38MAPK, #9212, Cell Signaling Technology, Danvers, MA, USA; 1:1,000), phospho-p38 MAPK (rabbit anti-Thr180/Tyr182, #9211, Cell Signaling Technology, Danvers, MA, USA; 1:1,000), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse anti-GAPDH, sc-47724, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:1,000).

    Techniques: Generated, Transmission Assay, Activation Assay, Inhibition

    Effect of luteolin on irradiation-induced interleukin-6 (IL-6) and matrix metalloproteinase-1 (MMP-1) expression ex vivo . Human skin explants were treated with luteolin (8 μg/mL), p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 (20 μM), or N -acetylcysteine (NAC; 816 μg/mL) before irradiation with a solar simulator (SS) at 6 J/cm 2 . At 24 hr after irradiation, the supernatant was collected. ( A ) Measurement with an enzyme-linked immunosorbent assay (ELISA) of IL-6 in the supernatant. ( B ) Measurement with an ELISA of MMP-1 in the supernatant. Values of irradiated skin explants were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±standard deviation (SD) of three independent experiments.

    Journal: Rejuvenation Research

    Article Title: Luteolin Prevents Solar Radiation-Induced Matrix Metalloproteinase-1 Activation in Human Fibroblasts: A Role for p38 Mitogen-Activated Protein Kinase and Interleukin-20 Released from Keratinocytes

    doi: 10.1089/rej.2011.1309

    Figure Lengend Snippet: Effect of luteolin on irradiation-induced interleukin-6 (IL-6) and matrix metalloproteinase-1 (MMP-1) expression ex vivo . Human skin explants were treated with luteolin (8 μg/mL), p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 (20 μM), or N -acetylcysteine (NAC; 816 μg/mL) before irradiation with a solar simulator (SS) at 6 J/cm 2 . At 24 hr after irradiation, the supernatant was collected. ( A ) Measurement with an enzyme-linked immunosorbent assay (ELISA) of IL-6 in the supernatant. ( B ) Measurement with an ELISA of MMP-1 in the supernatant. Values of irradiated skin explants were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±standard deviation (SD) of three independent experiments.

    Article Snippet: The following antibodies and dilutions were used for immunohistochemical staining or western blotting: Anti-p38 MAPK (Cell Signaling, Frankfurt, Germany), 1:1,000; anti-phospho-p38 MAPK (clone 3D7, Cell Signaling, Frankfurt, Germany); anti-HSC-70 (clone B-6, Santa Cruz Biotechnology, Heidelberg, Germany) 1:5,000; and neutralizing anti-IL-20 (RD Systems, Wiesbaden, Germany) 1:10.

    Techniques: Irradiation, Expressing, Ex Vivo, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Luteolin decreases ultraviolet A-1 (UVA-1)-induced interleukin-6 (IL-6), matrix metalloproteinase-1 (MMP-1) expression, and hyaluronidase activity in normal human dermal fibroblasts (NHDFs). NHDFs were incubated for 1 hr with luteolin (4 μg/mL) or p38 MAPK inhibitor SB203580 (20 μM) and exposed to 20 J/cm 2 UVA-1 radiation. At 24 hr after irradiation, the supernatant was analyzed. ( A ) Measurement of IL-6 with an enzyme-linked immunosorbent assay (ELISA) in the supernatant. ( B ) Measurement of MMP-1 expression and activity with an ELISA in the supernatant. Values of irradiated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. ( C ) Detection of hyaluronidase activity by hyaluronic acid (HA) zymography. The histograms depict relative protein activity levels to the irradiated sample. The values are means±standard deviation (SD) of three independent experiments (**) p

    Journal: Rejuvenation Research

    Article Title: Luteolin Prevents Solar Radiation-Induced Matrix Metalloproteinase-1 Activation in Human Fibroblasts: A Role for p38 Mitogen-Activated Protein Kinase and Interleukin-20 Released from Keratinocytes

    doi: 10.1089/rej.2011.1309

    Figure Lengend Snippet: Luteolin decreases ultraviolet A-1 (UVA-1)-induced interleukin-6 (IL-6), matrix metalloproteinase-1 (MMP-1) expression, and hyaluronidase activity in normal human dermal fibroblasts (NHDFs). NHDFs were incubated for 1 hr with luteolin (4 μg/mL) or p38 MAPK inhibitor SB203580 (20 μM) and exposed to 20 J/cm 2 UVA-1 radiation. At 24 hr after irradiation, the supernatant was analyzed. ( A ) Measurement of IL-6 with an enzyme-linked immunosorbent assay (ELISA) in the supernatant. ( B ) Measurement of MMP-1 expression and activity with an ELISA in the supernatant. Values of irradiated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. ( C ) Detection of hyaluronidase activity by hyaluronic acid (HA) zymography. The histograms depict relative protein activity levels to the irradiated sample. The values are means±standard deviation (SD) of three independent experiments (**) p

    Article Snippet: The following antibodies and dilutions were used for immunohistochemical staining or western blotting: Anti-p38 MAPK (Cell Signaling, Frankfurt, Germany), 1:1,000; anti-phospho-p38 MAPK (clone 3D7, Cell Signaling, Frankfurt, Germany); anti-HSC-70 (clone B-6, Santa Cruz Biotechnology, Heidelberg, Germany) 1:5,000; and neutralizing anti-IL-20 (RD Systems, Wiesbaden, Germany) 1:10.

    Techniques: Expressing, Activity Assay, Incubation, Irradiation, Enzyme-linked Immunosorbent Assay, Zymography, Standard Deviation

    Luteolin reduces ultraviolet A-1 (UVA-1)-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation in normal human dermal fibroblasts (NHDFs). NHDFs were treated for 30 min with different substances (4 μg/mL luteolin, 20 μM SB203580, 816 μg/ml NAC). Subsequently, the cells were irradiated with 20 J/cm 2 UVA-1. P38 MAPK, and phosphorylated p38 MAPK protein expression was assessed 30 min after irradiation by western blotting. The histogram depicting relative protein expression levels of phosphorylated p38 MAPK was normalized to the expression level of p38 MAPK. Values of irradiated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±(SD) of three independent experiments (**) p

    Journal: Rejuvenation Research

    Article Title: Luteolin Prevents Solar Radiation-Induced Matrix Metalloproteinase-1 Activation in Human Fibroblasts: A Role for p38 Mitogen-Activated Protein Kinase and Interleukin-20 Released from Keratinocytes

    doi: 10.1089/rej.2011.1309

    Figure Lengend Snippet: Luteolin reduces ultraviolet A-1 (UVA-1)-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation in normal human dermal fibroblasts (NHDFs). NHDFs were treated for 30 min with different substances (4 μg/mL luteolin, 20 μM SB203580, 816 μg/ml NAC). Subsequently, the cells were irradiated with 20 J/cm 2 UVA-1. P38 MAPK, and phosphorylated p38 MAPK protein expression was assessed 30 min after irradiation by western blotting. The histogram depicting relative protein expression levels of phosphorylated p38 MAPK was normalized to the expression level of p38 MAPK. Values of irradiated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±(SD) of three independent experiments (**) p

    Article Snippet: The following antibodies and dilutions were used for immunohistochemical staining or western blotting: Anti-p38 MAPK (Cell Signaling, Frankfurt, Germany), 1:1,000; anti-phospho-p38 MAPK (clone 3D7, Cell Signaling, Frankfurt, Germany); anti-HSC-70 (clone B-6, Santa Cruz Biotechnology, Heidelberg, Germany) 1:5,000; and neutralizing anti-IL-20 (RD Systems, Wiesbaden, Germany) 1:10.

    Techniques: Irradiation, Expressing, Western Blot

    Luteolin reduces solar simulator-induced interleukin-6 (IL-6) and IL-20 expression in keratinocytes and influences IL-20-induced matrix metalloproteinase-1 (MMP-1) expression and hyaluronidase activity in normal human dermal fibroblasts (NHDFs). HaCaT cells were treated for 1 hr with luteolin (8 μg/mL) or p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 (20 μM) and exposed to 6 J/cm 2 solar simulator (SS) irradiation on ice ( A–C ), and NHDFs were treated with IL-20 (10 ng/mL) with or without luteolin (4 μg/ml) for 30 min ( D–E ). After 24 hr, the culture supernatant was collected. ( A ) Amount of IL-6 protein in the culture supernatant of HaCaT cells 24 hr after irradiation was measured by enzyme-linked immunoassay (ELISA). Values of irradiated HaCaT cells were arbitrarily set to 100%; all other samples were indicated as percentage of control. ( B ) IL-20 gene expression was quantified by real-time quantitative PCR (RT-qPCR) in HaCaT cells. For each condition three RNAs isolated and transcribed independently were submitted in duplicate to the RT-qPCR reaction. Values of untreated HaCaT cells were arbitrarily set to 1; all other samples were indicated as fold change compared to control. ( C ) Amount of IL-20 protein in the culture supernatant of HaCaT cells 24 hr after irradiation as measured by ELISA. Values of irradiated HaCaT cells were arbitrarily set to 100%; all other samples were indicated as percentage of control. ( D ) Measurement of MMP-1 with an ELISA in the supernatant of NHDFs. ( E ) Detection of hyaluronidase activity by hyaluronic acid (HA) zymography in the supernatant of NHDFs. The histograms depict relative protein expression levels to the irradiated sample. Values of stimulated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±standard deviation (SD) of three independent experiments.

    Journal: Rejuvenation Research

    Article Title: Luteolin Prevents Solar Radiation-Induced Matrix Metalloproteinase-1 Activation in Human Fibroblasts: A Role for p38 Mitogen-Activated Protein Kinase and Interleukin-20 Released from Keratinocytes

    doi: 10.1089/rej.2011.1309

    Figure Lengend Snippet: Luteolin reduces solar simulator-induced interleukin-6 (IL-6) and IL-20 expression in keratinocytes and influences IL-20-induced matrix metalloproteinase-1 (MMP-1) expression and hyaluronidase activity in normal human dermal fibroblasts (NHDFs). HaCaT cells were treated for 1 hr with luteolin (8 μg/mL) or p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 (20 μM) and exposed to 6 J/cm 2 solar simulator (SS) irradiation on ice ( A–C ), and NHDFs were treated with IL-20 (10 ng/mL) with or without luteolin (4 μg/ml) for 30 min ( D–E ). After 24 hr, the culture supernatant was collected. ( A ) Amount of IL-6 protein in the culture supernatant of HaCaT cells 24 hr after irradiation was measured by enzyme-linked immunoassay (ELISA). Values of irradiated HaCaT cells were arbitrarily set to 100%; all other samples were indicated as percentage of control. ( B ) IL-20 gene expression was quantified by real-time quantitative PCR (RT-qPCR) in HaCaT cells. For each condition three RNAs isolated and transcribed independently were submitted in duplicate to the RT-qPCR reaction. Values of untreated HaCaT cells were arbitrarily set to 1; all other samples were indicated as fold change compared to control. ( C ) Amount of IL-20 protein in the culture supernatant of HaCaT cells 24 hr after irradiation as measured by ELISA. Values of irradiated HaCaT cells were arbitrarily set to 100%; all other samples were indicated as percentage of control. ( D ) Measurement of MMP-1 with an ELISA in the supernatant of NHDFs. ( E ) Detection of hyaluronidase activity by hyaluronic acid (HA) zymography in the supernatant of NHDFs. The histograms depict relative protein expression levels to the irradiated sample. Values of stimulated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±standard deviation (SD) of three independent experiments.

    Article Snippet: The following antibodies and dilutions were used for immunohistochemical staining or western blotting: Anti-p38 MAPK (Cell Signaling, Frankfurt, Germany), 1:1,000; anti-phospho-p38 MAPK (clone 3D7, Cell Signaling, Frankfurt, Germany); anti-HSC-70 (clone B-6, Santa Cruz Biotechnology, Heidelberg, Germany) 1:5,000; and neutralizing anti-IL-20 (RD Systems, Wiesbaden, Germany) 1:10.

    Techniques: Expressing, Activity Assay, Irradiation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Isolation, Zymography, Standard Deviation

    Conditioned medium of irradiated HaCaT cells influences interleukin-6 (IL-6) and matrix metalloproteinase-1 (MMP-1) expression in normal human dermal fibroblasts (NHDFs). In luteolin-pretreated HaCaT cells, this effect could be inhibited. HaCaT cells were treated as indicated under the graphic before irradiation (8 μg/mL luteolin, 20 μM p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580, 816 μg/mL N -acetylcysteine [NAC]). At 24 hr after irradiation at 6 J/cm 2 with a solar simulator, the conditioned medium was transferred to NHDF. The neutralizing IL-20 antibody was added after irradiation, as recommended by the manufacturer. ( A ) Measurement of IL-6 with an enzyme-linked immunosorbent assay (ELISA) in the NHDFs supernatant. ( B ) Measurement of MMP-1 with an ELISA in the NHDFs supernatant. ( C ) Detection of hyaluronidase activity by hyaluronic acid (HA) zymography. The active enzymes were detected as clear bands on a dark background after staining. The photo shows an inversion of the original gel to facilitate the detection of the bands. Values of irradiated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±standard deviation (SD) of three independent experiments.

    Journal: Rejuvenation Research

    Article Title: Luteolin Prevents Solar Radiation-Induced Matrix Metalloproteinase-1 Activation in Human Fibroblasts: A Role for p38 Mitogen-Activated Protein Kinase and Interleukin-20 Released from Keratinocytes

    doi: 10.1089/rej.2011.1309

    Figure Lengend Snippet: Conditioned medium of irradiated HaCaT cells influences interleukin-6 (IL-6) and matrix metalloproteinase-1 (MMP-1) expression in normal human dermal fibroblasts (NHDFs). In luteolin-pretreated HaCaT cells, this effect could be inhibited. HaCaT cells were treated as indicated under the graphic before irradiation (8 μg/mL luteolin, 20 μM p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580, 816 μg/mL N -acetylcysteine [NAC]). At 24 hr after irradiation at 6 J/cm 2 with a solar simulator, the conditioned medium was transferred to NHDF. The neutralizing IL-20 antibody was added after irradiation, as recommended by the manufacturer. ( A ) Measurement of IL-6 with an enzyme-linked immunosorbent assay (ELISA) in the NHDFs supernatant. ( B ) Measurement of MMP-1 with an ELISA in the NHDFs supernatant. ( C ) Detection of hyaluronidase activity by hyaluronic acid (HA) zymography. The active enzymes were detected as clear bands on a dark background after staining. The photo shows an inversion of the original gel to facilitate the detection of the bands. Values of irradiated NHDFs were arbitrarily set to 100%; all other samples were indicated as percentage of control. The values are means±standard deviation (SD) of three independent experiments.

    Article Snippet: The following antibodies and dilutions were used for immunohistochemical staining or western blotting: Anti-p38 MAPK (Cell Signaling, Frankfurt, Germany), 1:1,000; anti-phospho-p38 MAPK (clone 3D7, Cell Signaling, Frankfurt, Germany); anti-HSC-70 (clone B-6, Santa Cruz Biotechnology, Heidelberg, Germany) 1:5,000; and neutralizing anti-IL-20 (RD Systems, Wiesbaden, Germany) 1:10.

    Techniques: Irradiation, Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay, Zymography, Staining, Standard Deviation

    Effects of coal-fired PM 2.5 on MAPK signaling pathways ( a ) p-p38, ( b ) p-JNK, and ( c ) p-ERK (tubulin was considered as an internal control; compared with HFD control mice, * P

    Journal: BMC Pharmacology & Toxicology

    Article Title: Effects of coal-fired PM2.5 on the expression levels of atherosclerosis-related proteins and the phosphorylation level of MAPK in ApoE−/− mice

    doi: 10.1186/s40360-020-00411-8

    Figure Lengend Snippet: Effects of coal-fired PM 2.5 on MAPK signaling pathways ( a ) p-p38, ( b ) p-JNK, and ( c ) p-ERK (tubulin was considered as an internal control; compared with HFD control mice, * P

    Article Snippet: Antibodies for p-p38 (4511S), p38 (9212S), p-JNK (4668S), JNK (9252S), p-ERK (9101S), ERK (9102S), and β-Tubulin (2146S) were obtained from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Mouse Assay

    Proposed mechanism by which short-term TAK1 inhibition protects neurons against ischemia. Acute TAK1 inhibition by OZ blocks activation of JNK, p38/MAPK, and NF- κ B, and thereby provides neuroprotection. Chronic TAK1 inhibition induces expression of ASK-1 that compensates for TAK1 inhibition with respect to JNK and p38/MAPK activation, transcription of Cox-2 , p40 phox , and Nox-2 and formation of reactive oxygen species, paving the way to cell death

    Journal: Cell Death and Differentiation

    Article Title: Acute inhibition of TAK1 protects against neuronal death in cerebral ischemia

    doi: 10.1038/cdd.2011.29

    Figure Lengend Snippet: Proposed mechanism by which short-term TAK1 inhibition protects neurons against ischemia. Acute TAK1 inhibition by OZ blocks activation of JNK, p38/MAPK, and NF- κ B, and thereby provides neuroprotection. Chronic TAK1 inhibition induces expression of ASK-1 that compensates for TAK1 inhibition with respect to JNK and p38/MAPK activation, transcription of Cox-2 , p40 phox , and Nox-2 and formation of reactive oxygen species, paving the way to cell death

    Article Snippet: Phospho-c-Jun, c-Jun, phospho-p38/MAPK, and p38/MAPK were detected with rabbit anti-phospho-c-Jun and anti-c-Jun, anti-phospho-p38/MAPK (Thr 180/Tyr 182), and anti-p38/MAPK antibodies (Cell Signaling, Danvers, MA, USA).

    Techniques: Inhibition, Activation Assay, Expressing

    Transient effect of TAK1 inhibition on p38/MAPK and JNK but not on NF- κ B signaling in vitro . ( a – c ) After incubation for 10 days or 1 h with OZ (600 nM) or DMSO, neurons were exposed to OGD followed by increasing recovery times. Controls received no OGD (Ø). Protein lysates were analyzed by immunoblotting to determine the degradation of I κ B α , a marker of NF- κ B activation, and the phosphorylation of p38/MAPK and c-Jun. Values are means±S.E.M., of four independent experiments. *** P

    Journal: Cell Death and Differentiation

    Article Title: Acute inhibition of TAK1 protects against neuronal death in cerebral ischemia

    doi: 10.1038/cdd.2011.29

    Figure Lengend Snippet: Transient effect of TAK1 inhibition on p38/MAPK and JNK but not on NF- κ B signaling in vitro . ( a – c ) After incubation for 10 days or 1 h with OZ (600 nM) or DMSO, neurons were exposed to OGD followed by increasing recovery times. Controls received no OGD (Ø). Protein lysates were analyzed by immunoblotting to determine the degradation of I κ B α , a marker of NF- κ B activation, and the phosphorylation of p38/MAPK and c-Jun. Values are means±S.E.M., of four independent experiments. *** P

    Article Snippet: Phospho-c-Jun, c-Jun, phospho-p38/MAPK, and p38/MAPK were detected with rabbit anti-phospho-c-Jun and anti-c-Jun, anti-phospho-p38/MAPK (Thr 180/Tyr 182), and anti-p38/MAPK antibodies (Cell Signaling, Danvers, MA, USA).

    Techniques: Inhibition, In Vitro, Incubation, Marker, Activation Assay

    Nck1, but not Nck2, is essential for activation of Erk1/2 and MEK1/2 proteins. Cells from each monoclonal group were untreated or treated with soluble CD3 antibody (1 μg/ml) at various time points. Cell lysates were subjected to immunoblot with anti-phospho-Erk1/2 antibody (A) . Below, the quantified signal intensity of the pErk1/2 was normalized to its total kinase and this value was relative to that in the unstimulated control cells (Mock), set as 1 (black dashed line) and plotted in bar graph. B) Immunoblot analysis of phospho-MEK1/2 from the lysates of cells that were stimulated as in A . Below, signal intensity was quantified and presented as a ratio of p-MEK1/2 to actin relative to that in unstimulated control cells (Mock), set as 1 (black dashed line). C) Immunoblot analysis of phospho-p38. Below, signal intensity was quantified and presented as described in A . D) . Polyclonal cells from Mock, Nck1- and Nck2-knockdown cells were stimulated as in A . Below, signal intensity was quantified and presented as described in A . E) Both Nck1 and Nck2 genes were co-silenced in Jurkat T cells. Nck expression levels were analysed by immunoblotting using indicated antibodies. F) The expression levels of surface CD3 molecules on double knockdown Nck1/2 were measured by flow cytometry. G) Cells from each monoclonal group, Nck1-, Nck2- and Nck1/2-knockdown cells were treated as in A for 0, 3 and 30 min. Lysates were subjected to immunoblot with anti-phospho-Erk1/2 antibody. Below, signal intensity was quantified and presented as described in A . Data are representative of at least two independent experiments.

    Journal: Cell Communication and Signaling : CCS

    Article Title: Non-overlapping functions of Nck1 and Nck2 adaptor proteins in T cell activation

    doi: 10.1186/1478-811X-12-21

    Figure Lengend Snippet: Nck1, but not Nck2, is essential for activation of Erk1/2 and MEK1/2 proteins. Cells from each monoclonal group were untreated or treated with soluble CD3 antibody (1 μg/ml) at various time points. Cell lysates were subjected to immunoblot with anti-phospho-Erk1/2 antibody (A) . Below, the quantified signal intensity of the pErk1/2 was normalized to its total kinase and this value was relative to that in the unstimulated control cells (Mock), set as 1 (black dashed line) and plotted in bar graph. B) Immunoblot analysis of phospho-MEK1/2 from the lysates of cells that were stimulated as in A . Below, signal intensity was quantified and presented as a ratio of p-MEK1/2 to actin relative to that in unstimulated control cells (Mock), set as 1 (black dashed line). C) Immunoblot analysis of phospho-p38. Below, signal intensity was quantified and presented as described in A . D) . Polyclonal cells from Mock, Nck1- and Nck2-knockdown cells were stimulated as in A . Below, signal intensity was quantified and presented as described in A . E) Both Nck1 and Nck2 genes were co-silenced in Jurkat T cells. Nck expression levels were analysed by immunoblotting using indicated antibodies. F) The expression levels of surface CD3 molecules on double knockdown Nck1/2 were measured by flow cytometry. G) Cells from each monoclonal group, Nck1-, Nck2- and Nck1/2-knockdown cells were treated as in A for 0, 3 and 30 min. Lysates were subjected to immunoblot with anti-phospho-Erk1/2 antibody. Below, signal intensity was quantified and presented as described in A . Data are representative of at least two independent experiments.

    Article Snippet: Anti-Nck1, anti-phospho-MEK1/2 and anti-phospho-p38, antibodies were from Cell Signaling (Cell Signaling Technology, Danver, MA, USA).

    Techniques: Activation Assay, Expressing, Flow Cytometry, Cytometry

    K1016A substitution in the DEP-1 KIM-like motif strongly impairs ERK1/2 recruitment. A , alignment of ERK1/2 KIM domains. These sites share an N-terminal hydrophobic residue (Ø H ), one or two positively charged residues ( basic ), and the ØA X ). B , HEK293 cells were transiently transfected with full-length K1016A DEP-1 mutant and with the wild type ( WT ) phosphatase. After 5 min of EGF induction, cells were lysed and the transfection efficiency monitored by immunoblotting with anti-DEP-1 and anti-tubulin antibodies. The phosphorylation levels of the ERK and p38 MAPKs were monitored by probing the protein extracts with anti-phospho-ERK1/2 ( WB: α-ERK1/2 ( P )), anti-ERK1/2 ( WB: α-ERK1/2 ), anti-phospho-p38 ( WB: α-p38 ( P )), and anti-p38 ( WB: α-p38 ) antibodies. NT , not transfected. C , whole protein extracts were immunoprecipitated with anti-HA antibody to recover DEP-1 protein. The immunoprecipitated sample ( IP ) was probed with anti-HA ( WB: α-HA ), anti-phospho-ERK1/2 ( WB: α-ERK1/2 (P)), anti-ERK1/2 ( WB: α-ERK1/2 ), and anti-p38 ( WB: α-p38 ). D , HEK293 cells were transiently co-transfected with full-length wild type phosphatase or its mutant K1016A and 3 μg of green fluorescent protein expression plasmid. After fixation, cells were stained with anti-DEP-1 antibody and analyzed by indirect immunofluorescence microscopy.

    Journal: The Journal of Biological Chemistry

    Article Title: Tumor Suppressor Density-enhanced Phosphatase-1 (DEP-1) Inhibits the RAS Pathway by Direct Dephosphorylation of ERK1/2 Kinases *

    doi: 10.1074/jbc.M109.002758

    Figure Lengend Snippet: K1016A substitution in the DEP-1 KIM-like motif strongly impairs ERK1/2 recruitment. A , alignment of ERK1/2 KIM domains. These sites share an N-terminal hydrophobic residue (Ø H ), one or two positively charged residues ( basic ), and the ØA X ). B , HEK293 cells were transiently transfected with full-length K1016A DEP-1 mutant and with the wild type ( WT ) phosphatase. After 5 min of EGF induction, cells were lysed and the transfection efficiency monitored by immunoblotting with anti-DEP-1 and anti-tubulin antibodies. The phosphorylation levels of the ERK and p38 MAPKs were monitored by probing the protein extracts with anti-phospho-ERK1/2 ( WB: α-ERK1/2 ( P )), anti-ERK1/2 ( WB: α-ERK1/2 ), anti-phospho-p38 ( WB: α-p38 ( P )), and anti-p38 ( WB: α-p38 ) antibodies. NT , not transfected. C , whole protein extracts were immunoprecipitated with anti-HA antibody to recover DEP-1 protein. The immunoprecipitated sample ( IP ) was probed with anti-HA ( WB: α-HA ), anti-phospho-ERK1/2 ( WB: α-ERK1/2 (P)), anti-ERK1/2 ( WB: α-ERK1/2 ), and anti-p38 ( WB: α-p38 ). D , HEK293 cells were transiently co-transfected with full-length wild type phosphatase or its mutant K1016A and 3 μg of green fluorescent protein expression plasmid. After fixation, cells were stained with anti-DEP-1 antibody and analyzed by indirect immunofluorescence microscopy.

    Article Snippet: Anti-hemagglutinin (HA) and anti-FLAG were from Sigma; anti-DEP-1, anti-SRC, and anti-tubulin were from Santa Cruz Biotechnology; anti-ERK1/2(P), anti-ERK1/2, anti-MEK(P), anti-MEK, anti-p38(P) and anti-p38 were from Cell Signaling, and anti-4G10 was from Upstate Biotechnology, Inc. Peroxidase-conjugated anti-rabbit, anti-mouse secondary antibodies were from Jackson ImmunoResearch, and anti-mouse TRICT was purchased from Molecular Probes.

    Techniques: Transfection, Mutagenesis, Western Blot, Immunoprecipitation, Expressing, Plasmid Preparation, Staining, Immunofluorescence, Microscopy

    p38 and JNK phosphorylation in the CPu in D1, D3 receptor mutant, and wild-type mice is not obviously changed after acute cocaine treatment. Time course of cocaine-induced phosphorylation of p38 and JNK in the CPu in wild-type ( A ), D1 ( B ), and D3 receptor mutant ( C ) mice. The phosphorylation status of p38 and JNK was determined 10, 20, 60, and 120 min after a cocaine injection (30 mg/kg, i.p.). Total cell extracts were analyzed by Western blotting using antibodies against dually phosphorylated-p38 (Thr 180 and Tyr 182 ), dually phosphorylated-JNK (Thr 183 and Tyr 185 ), total p38, or JNK, respectively. The same set of mice used in probing ERK activation was used in analyzing p38 and JNK activation after cocaine injections.

    Journal: The Journal of Neuroscience

    Article Title: Cocaine-Induced Intracellular Signaling and Gene Expression Are Oppositely Regulated by the Dopamine D1 and D3 Receptors

    doi: 10.1523/JNEUROSCI.0060-04.2004

    Figure Lengend Snippet: p38 and JNK phosphorylation in the CPu in D1, D3 receptor mutant, and wild-type mice is not obviously changed after acute cocaine treatment. Time course of cocaine-induced phosphorylation of p38 and JNK in the CPu in wild-type ( A ), D1 ( B ), and D3 receptor mutant ( C ) mice. The phosphorylation status of p38 and JNK was determined 10, 20, 60, and 120 min after a cocaine injection (30 mg/kg, i.p.). Total cell extracts were analyzed by Western blotting using antibodies against dually phosphorylated-p38 (Thr 180 and Tyr 182 ), dually phosphorylated-JNK (Thr 183 and Tyr 185 ), total p38, or JNK, respectively. The same set of mice used in probing ERK activation was used in analyzing p38 and JNK activation after cocaine injections.

    Article Snippet: Antibodies against ERK, JNK, p38 and phospho-ERK, phospho-JNK, phospho-p38 (Cell Signaling Technology, Beverly, MA) and c-Fos, neogenin and synaptotagmin VII (Santa Cruz Biotechnology, Santa Cruz, CA) were used at 1:1000 dilution, and antibodies against actin (Santa Cruz Biotechnology) were used at the 1:3000 dilution.

    Techniques: Mutagenesis, Mouse Assay, Injection, Western Blot, Activation Assay

    Activation of p38 MAPK is necessary for the phagocytic activity of microglia. A , Western blot analysis for various MAPKs. Co-cultures of cortical explants and MG5 cells were incubated for 12 h, and neurites were then transected. Degenerating axons and

    Journal:

    Article Title: Engulfment of Axon Debris by Microglia Requires p38 MAPK Activity *

    doi: 10.1074/jbc.M109.005603

    Figure Lengend Snippet: Activation of p38 MAPK is necessary for the phagocytic activity of microglia. A , Western blot analysis for various MAPKs. Co-cultures of cortical explants and MG5 cells were incubated for 12 h, and neurites were then transected. Degenerating axons and

    Article Snippet: Cells were fixed in 4% paraformaldehyde in 0.1 m phosphate buffer for 1 h at room temperature and incubated with a blocking solution containing 1% bovine serum albumin and 0.1% Triton X-100 in phosphate-buffered saline for 1 h, followed by overnight incubation at 4 °C with anti-β-tubulin III (Tuj-1) antibody (diluted 1:1000 in the blocking solution; Covance, Princeton, NJ), anti-ED1 (CD68) antibody (diluted 1:100 in the blocking solution; AbD Serotec, Raleigh, NC), anti-GAP43 antibody (diluted 1:1000 in the blocking solution; Chemicon/Millipore, Temecula, CA), and anti-phospho-p38 antibody (diluted 1:100 in the blocking solution; Cell Signaling Technology, Beverly, MA).

    Techniques: Activation Assay, Activity Assay, Western Blot, Incubation

    p38 and JNK MAPK pathways regulate OCP crystal induced NO production and iNOS mRNA expression. Nonadherent bovine articular chondrocytes were stimulated with OCP crystals or IL-1β for 24 hours with or without pretreatment (1 hour) with (a,c) SB 203580 (a p38 MAPK inhibitor), (b,c) JNK II inhibitor or (d) PD 58059 (an Erk1/2 MAPK inhibitor). iNOS transcripts (panel d) were assessed 8 hours after stimulation by OCP crystals or IL-1β by RT-PCR. The RT-PCR was representative of three experiments. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; iNOS, inducible nitric oxide synthase; JNK, c-Jun amino-terminal kinase; NO, nitric oxide; OCP, octacalcium phosphate.

    Journal: Arthritis Research & Therapy

    Article Title: Octacalcium phosphate crystals directly stimulate expression of inducible nitric oxide synthase through p38 and JNK mitogen-activated protein kinases in articular chondrocytes

    doi: 10.1186/ar1763

    Figure Lengend Snippet: p38 and JNK MAPK pathways regulate OCP crystal induced NO production and iNOS mRNA expression. Nonadherent bovine articular chondrocytes were stimulated with OCP crystals or IL-1β for 24 hours with or without pretreatment (1 hour) with (a,c) SB 203580 (a p38 MAPK inhibitor), (b,c) JNK II inhibitor or (d) PD 58059 (an Erk1/2 MAPK inhibitor). iNOS transcripts (panel d) were assessed 8 hours after stimulation by OCP crystals or IL-1β by RT-PCR. The RT-PCR was representative of three experiments. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; iNOS, inducible nitric oxide synthase; JNK, c-Jun amino-terminal kinase; NO, nitric oxide; OCP, octacalcium phosphate.

    Article Snippet: Antibodies Phospho-specific JNK (Thr183/Tyr185) and p38 (Thr180/Tyr182), and total JNK and p38 antibodies were purchased from Cell Signaling Technology (Ozyme, St Quentin Yvelines, France).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    OCP crystal induced p38 and JNK MAPK activation. Nonadherent bovine articular chondrocytes were stimulated with OCP crystals for 20 min with or without pretreatment (1 hour) with SB 203580 or JNK II inhibitor. Fifteen (p38 MAPK) or 30 μg (JNK MAPK) protein of cell lysates were subjected to SDS-PAGE/Western blot analysis. The blots were probed with phosphospecific (a) p38 and (b) JNK MAPK antibodies. To ensure equal loading, the blot was stripped and reprobed with total p38 MAPK or JNK antibodies. JNK, c-Jun amino-terminal kinase; OCP, octacalcium phosphate.

    Journal: Arthritis Research & Therapy

    Article Title: Octacalcium phosphate crystals directly stimulate expression of inducible nitric oxide synthase through p38 and JNK mitogen-activated protein kinases in articular chondrocytes

    doi: 10.1186/ar1763

    Figure Lengend Snippet: OCP crystal induced p38 and JNK MAPK activation. Nonadherent bovine articular chondrocytes were stimulated with OCP crystals for 20 min with or without pretreatment (1 hour) with SB 203580 or JNK II inhibitor. Fifteen (p38 MAPK) or 30 μg (JNK MAPK) protein of cell lysates were subjected to SDS-PAGE/Western blot analysis. The blots were probed with phosphospecific (a) p38 and (b) JNK MAPK antibodies. To ensure equal loading, the blot was stripped and reprobed with total p38 MAPK or JNK antibodies. JNK, c-Jun amino-terminal kinase; OCP, octacalcium phosphate.

    Article Snippet: Antibodies Phospho-specific JNK (Thr183/Tyr185) and p38 (Thr180/Tyr182), and total JNK and p38 antibodies were purchased from Cell Signaling Technology (Ozyme, St Quentin Yvelines, France).

    Techniques: Activation Assay, SDS Page, Western Blot

    Fibronectin synthesis in HK-2 cells by ELISA. Fibronectin was measured by ELISA of the HK-2 cells in 5.5 mM glucose (lane 1), 30 mM glucose(lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5) and 30 mM glucose+AP-1 inhibitor(lane 6). Values represent the mean ± SD, a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Fibronectin synthesis in HK-2 cells by ELISA. Fibronectin was measured by ELISA of the HK-2 cells in 5.5 mM glucose (lane 1), 30 mM glucose(lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5) and 30 mM glucose+AP-1 inhibitor(lane 6). Values represent the mean ± SD, a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Enzyme-linked Immunosorbent Assay

    Expression and phosphorylation of p38 MAPK in HK-2 cells by Western blot. Expression and phosphorylation of p38 MAPK under 5.5 mM glucose (lane 1), 30 mM glucose(lane 2) and 30 mM glucose+AP-1 inhibitor were determined by. a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Expression and phosphorylation of p38 MAPK in HK-2 cells by Western blot. Expression and phosphorylation of p38 MAPK under 5.5 mM glucose (lane 1), 30 mM glucose(lane 2) and 30 mM glucose+AP-1 inhibitor were determined by. a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Expressing, Western Blot

    Activity of AP-1 in HK-2 cells by EMSA. (A)AP-1 activity was analyzed by EMSA of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose(lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h (lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5).(B) AP-1 activity was analyzed by EMSA of the HK-2 cells cultured in 5.5 mM glucose(lane 1), 30 mM glucose (lane 2) and 30 mM glucose+AP-1 inhibitor(lane 3). Values represent the mean ± SD, a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Activity of AP-1 in HK-2 cells by EMSA. (A)AP-1 activity was analyzed by EMSA of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose(lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h (lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5).(B) AP-1 activity was analyzed by EMSA of the HK-2 cells cultured in 5.5 mM glucose(lane 1), 30 mM glucose (lane 2) and 30 mM glucose+AP-1 inhibitor(lane 3). Values represent the mean ± SD, a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Activity Assay, Cell Culture

    Expression of E-cadherin, CK, α-SMA, vimentin in HK-2 cells by Western blot and RT-PCR analysis. (A–B)E-cadherin expression(A1,A2) and α-SMA(B1,B2) were analyzed by Western blot of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5).(C–D)mRNA of CK (C1,C2) and vimentin(D1,D2) were analyzed by RT-PCR of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5).Values represent the mean ± SD, a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Expression of E-cadherin, CK, α-SMA, vimentin in HK-2 cells by Western blot and RT-PCR analysis. (A–B)E-cadherin expression(A1,A2) and α-SMA(B1,B2) were analyzed by Western blot of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5).(C–D)mRNA of CK (C1,C2) and vimentin(D1,D2) were analyzed by RT-PCR of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5).Values represent the mean ± SD, a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Cell Culture

    Expression of E-cadherin, CK, α-SMA, vimentin in HK-2 cells by Western blot. (A)α-SMA, (B)E-caderin, (C)CK, (D)vimentin were measured by Western blot of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 5.5 mM glucose with p38 siRNA (lane 2), 5.5 mM glucose with AP-1 inhibitor (lane 3), 5.5 mM glucose+TGF-β1 (10 ng/ml) (lane 4), 5.5 mM glucose+TGF-β1(10 ng/ml) with p38 siRNA (lane 5), 5.5 mM glucose+TGF-β1(10 ng/ml) with the AP-1 inhibitor (lane 6). a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Expression of E-cadherin, CK, α-SMA, vimentin in HK-2 cells by Western blot. (A)α-SMA, (B)E-caderin, (C)CK, (D)vimentin were measured by Western blot of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 5.5 mM glucose with p38 siRNA (lane 2), 5.5 mM glucose with AP-1 inhibitor (lane 3), 5.5 mM glucose+TGF-β1 (10 ng/ml) (lane 4), 5.5 mM glucose+TGF-β1(10 ng/ml) with p38 siRNA (lane 5), 5.5 mM glucose+TGF-β1(10 ng/ml) with the AP-1 inhibitor (lane 6). a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Expressing, Western Blot, Cell Culture

    The expression of Snail mRNA in HK-2 cells by RT-PCR. (A)The levels of Snail mRNA were detected by RT-PCR of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA (lane 3), 30 mM glucose+p38 siRNA for 24 h (lane 4), 30 mM glucose DMEM+p38 siRNA for 48 h(lane 5).(B)The levels of the Snail mRNA were detected by RT-PCR of the HK-2 cells incubated in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), and 30 mM glucose+AP-1 inhibitor (lane 3). Values represent the mean ± SD, aP

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: The expression of Snail mRNA in HK-2 cells by RT-PCR. (A)The levels of Snail mRNA were detected by RT-PCR of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA (lane 3), 30 mM glucose+p38 siRNA for 24 h (lane 4), 30 mM glucose DMEM+p38 siRNA for 48 h(lane 5).(B)The levels of the Snail mRNA were detected by RT-PCR of the HK-2 cells incubated in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), and 30 mM glucose+AP-1 inhibitor (lane 3). Values represent the mean ± SD, aP

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Incubation

    Transmission electron microscopic analysis of the HK-2 cells cultured under different conditions. (A–C)The HK-2 cells under 30 mM glucose (B) or 30 mM glucose+Cont siRNA (C) showed decreased number of microvilli, mitochondria and increased volume density of rough endoplasmic reticulum compared to that of the cells cultured in 5.5 mM glucose(A). (D–F)Changes of the cells were reversed,which were cultured in the 30 mM glucose+p38 siRNA for 24 h(D) or 48 h(E) or 30 mM glucose+AP-1 inhibitor(F).

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Transmission electron microscopic analysis of the HK-2 cells cultured under different conditions. (A–C)The HK-2 cells under 30 mM glucose (B) or 30 mM glucose+Cont siRNA (C) showed decreased number of microvilli, mitochondria and increased volume density of rough endoplasmic reticulum compared to that of the cells cultured in 5.5 mM glucose(A). (D–F)Changes of the cells were reversed,which were cultured in the 30 mM glucose+p38 siRNA for 24 h(D) or 48 h(E) or 30 mM glucose+AP-1 inhibitor(F).

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Transmission Assay, Cell Culture

    The expression of p38 MAPK and phosphorylated P38 MAPK by Western blot analysis. p38 MAPK and phosphorylated p38 MAPK were measured by Western blot of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5). Values represent the mean ± SD, a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: The expression of p38 MAPK and phosphorylated P38 MAPK by Western blot analysis. p38 MAPK and phosphorylated p38 MAPK were measured by Western blot of the HK-2 cells cultured in 5.5 mM glucose (lane 1), 30 mM glucose (lane 2), 30 mM glucose+Cont siRNA(lane 3), 30 mM glucose+p38 siRNA for 24 h(lane 4), 30 mM glucose+p38 siRNA for 48 h(lane 5). Values represent the mean ± SD, a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Expressing, Western Blot, Cell Culture

    Effect of knockdown of p38MAPK by p38 MAPK siRNA on p38 MAPK expression by Western blot and RT-PCR analysis. (A)P38 MAPK expression were determined by RT-PCR of the HK-2 cells cultured for 72 h in 5.5 mM glucose (lane 1), 5.5 mM glucose+Cont siRNA (lane 2), 5.5 mM glucose+p38 siRNA (lane 3). (B)P38 MAPK expression were determined by Western blot of the HK-2 cells cultured for 72 h in 5.5 mM glucose (lane 1), 5.5 mM glucose+Cont siRNA (lane 2), 5.5 mM glucose+p38 siRNA (lane 3).Values represent the mean ± SD, a P

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Effect of knockdown of p38MAPK by p38 MAPK siRNA on p38 MAPK expression by Western blot and RT-PCR analysis. (A)P38 MAPK expression were determined by RT-PCR of the HK-2 cells cultured for 72 h in 5.5 mM glucose (lane 1), 5.5 mM glucose+Cont siRNA (lane 2), 5.5 mM glucose+p38 siRNA (lane 3). (B)P38 MAPK expression were determined by Western blot of the HK-2 cells cultured for 72 h in 5.5 mM glucose (lane 1), 5.5 mM glucose+Cont siRNA (lane 2), 5.5 mM glucose+p38 siRNA (lane 3).Values represent the mean ± SD, a P

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Cell Culture

    Inverted microscope analysis of the HK-2 cells cultured under different conditions. (A) A typical epithelial cuboidal shape of the HK-2 cells cultured in 5.5 mM glucose condition were shown, with the characteristic cobblestone morphology. (B–C)Morphological changes of the HK-2 cells. The cells became more elongated, less adhered and lost their apical-to-basal polarity after treated with 30 mM glucose(B) or 30 mM glucose+Cont siRNA(C).(D–F)Changes of the cells were reversed, which were exposed to 30 mM glucose+p38 siRNA for 24 h(D) or for 48 h(E) or 30 mM glucose+AP-1 inhibitor(F).

    Journal: PLoS ONE

    Article Title: The Role of the p38 MAPK Signaling Pathway in High Glucose-Induced Epithelial-Mesenchymal Transition of Cultured Human Renal Tubular Epithelial Cells

    doi: 10.1371/journal.pone.0022806

    Figure Lengend Snippet: Inverted microscope analysis of the HK-2 cells cultured under different conditions. (A) A typical epithelial cuboidal shape of the HK-2 cells cultured in 5.5 mM glucose condition were shown, with the characteristic cobblestone morphology. (B–C)Morphological changes of the HK-2 cells. The cells became more elongated, less adhered and lost their apical-to-basal polarity after treated with 30 mM glucose(B) or 30 mM glucose+Cont siRNA(C).(D–F)Changes of the cells were reversed, which were exposed to 30 mM glucose+p38 siRNA for 24 h(D) or for 48 h(E) or 30 mM glucose+AP-1 inhibitor(F).

    Article Snippet: Antibodies and chemical inhibitors Anti-phospho-p38 antibodies and anti-p38 antibodies were purchased from Cell Signaling Technology (USA).

    Techniques: Inverted Microscopy, Cell Culture

    Lpcat3 KO macrophages promotes p38, p42/44, and NFκB activation. WT and Lpcat3 KO macrophages were treated with 1 μg/ml LPS. (A) Western blot and quantitative display of total p38 and phosphorylated p38. (B) Western blot and quantitative display of total p42/44 and phosphorylated p42/44. Macrophage nucleus were isolated and p65 was measured by Western blot. (C) Western blot and quantitative display of p65. Values are mean ± SD, n = 3–4, * P

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Macrophage Lysophosphatidylcholine Acyltransferase 3 Deficiency-Mediated Inflammation Is Not Sufficient to Induce Atherosclerosis in a Mouse Model

    doi: 10.3389/fcvm.2018.00192

    Figure Lengend Snippet: Lpcat3 KO macrophages promotes p38, p42/44, and NFκB activation. WT and Lpcat3 KO macrophages were treated with 1 μg/ml LPS. (A) Western blot and quantitative display of total p38 and phosphorylated p38. (B) Western blot and quantitative display of total p42/44 and phosphorylated p42/44. Macrophage nucleus were isolated and p65 was measured by Western blot. (C) Western blot and quantitative display of p65. Values are mean ± SD, n = 3–4, * P

    Article Snippet: Cell lysates were used for Western blots with antibodies against p38 and p42/44 (Cell Signaling).

    Techniques: Activation Assay, Western Blot, Isolation

    The relative mRNA expressions of STE20, TRAF6, IGF1R, and p38 MAPK were shown at the 3 th , 7 th , 14 th , and 21 th day. The data are expressed as mean ± SD. ∗P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Effect of Plastrum Testudinis Extracts on the Proliferation and Osteogenic Differentiation of rBMSCs by Regulating p38 MAPK-Related Genes

    doi: 10.1155/2019/6815620

    Figure Lengend Snippet: The relative mRNA expressions of STE20, TRAF6, IGF1R, and p38 MAPK were shown at the 3 th , 7 th , 14 th , and 21 th day. The data are expressed as mean ± SD. ∗P

    Article Snippet: After blocking in 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4°C with antibodies specific to GAPDH (RC-5G5; 1:10000; KangChen Bio-tech, Shanghai, China), TRAF6 (ab33915; 1:8000; Abcam, Cambridge, UK), p-p38 MAPK (#4511; 1:1000; CST, Boston, USA), STE20 (ab155198; 1:3000; Abcam, Cambridge, UK), and IGF1R (ab79176; 1:2000; Abcam, Cambridge, UK).

    Techniques:

    Protein expression levels of TRAF6, STE20, IGF1R, and p-p38 MAPK were analysed by Western Blot. The data are expressed as mean ± SD. ∗ P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Effect of Plastrum Testudinis Extracts on the Proliferation and Osteogenic Differentiation of rBMSCs by Regulating p38 MAPK-Related Genes

    doi: 10.1155/2019/6815620

    Figure Lengend Snippet: Protein expression levels of TRAF6, STE20, IGF1R, and p-p38 MAPK were analysed by Western Blot. The data are expressed as mean ± SD. ∗ P

    Article Snippet: After blocking in 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4°C with antibodies specific to GAPDH (RC-5G5; 1:10000; KangChen Bio-tech, Shanghai, China), TRAF6 (ab33915; 1:8000; Abcam, Cambridge, UK), p-p38 MAPK (#4511; 1:1000; CST, Boston, USA), STE20 (ab155198; 1:3000; Abcam, Cambridge, UK), and IGF1R (ab79176; 1:2000; Abcam, Cambridge, UK).

    Techniques: Expressing, Western Blot

    Schematic of PTE on proliferation and osteogenic differentiation of rBMSCs by regulating p38 MAPK-related genes based on our findings.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Effect of Plastrum Testudinis Extracts on the Proliferation and Osteogenic Differentiation of rBMSCs by Regulating p38 MAPK-Related Genes

    doi: 10.1155/2019/6815620

    Figure Lengend Snippet: Schematic of PTE on proliferation and osteogenic differentiation of rBMSCs by regulating p38 MAPK-related genes based on our findings.

    Article Snippet: After blocking in 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4°C with antibodies specific to GAPDH (RC-5G5; 1:10000; KangChen Bio-tech, Shanghai, China), TRAF6 (ab33915; 1:8000; Abcam, Cambridge, UK), p-p38 MAPK (#4511; 1:1000; CST, Boston, USA), STE20 (ab155198; 1:3000; Abcam, Cambridge, UK), and IGF1R (ab79176; 1:2000; Abcam, Cambridge, UK).

    Techniques:

    The relative mRNA expressions of STE20, TRAF6, IGF1R, and p38 MAPK were shown at the 3 th , 7 th , 14 th , and 21 th day. The data are expressed as mean ± SD. ∗P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Effect of Plastrum Testudinis Extracts on the Proliferation and Osteogenic Differentiation of rBMSCs by Regulating p38 MAPK-Related Genes

    doi: 10.1155/2019/6815620

    Figure Lengend Snippet: The relative mRNA expressions of STE20, TRAF6, IGF1R, and p38 MAPK were shown at the 3 th , 7 th , 14 th , and 21 th day. The data are expressed as mean ± SD. ∗P

    Article Snippet: After blocking in 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4°C with antibodies specific to GAPDH (RC-5G5; 1:10000; KangChen Bio-tech, Shanghai, China), TRAF6 (ab33915; 1:8000; Abcam, Cambridge, UK), p-p38 MAPK (#4511; 1:1000; CST, Boston, USA), STE20 (ab155198; 1:3000; Abcam, Cambridge, UK), and IGF1R (ab79176; 1:2000; Abcam, Cambridge, UK).

    Techniques:

    Calcium ionophore stimulation results in p38 MAPK phosphorylation in C2C12 myotubes. To determine whether calcium ionophore treatment resulted in activation/phosphorylation of p38 MAPK, myotubes were treated for 3 h with 1 μM A23187 or control, and protein was isolated 30 min and 24 h following treatment. Calcium ionophore treatment resulted in significant phosphorylation of p38 MAPK compared with BL. Densitometric analysis ( top ) and representative immunoblot images ( bottom ) are shown. Values are means ± SE. Phosphorylated-to-total protein content ratios were derived and then expressed relative to the BL group for comparisons. GAPDH was a loading reference. BL, vehicle treated; A23187, treated for 3 h, and protein isolated 30 min posttreatment; 24H post, myotubes treated for 3 h, protein isolated 24 h posttreatment. All experiments were performed in duplicate on at least 3 separate occasions, in serial passage. *A23187 significantly different than BL ( P

    Journal: Journal of Applied Physiology

    Article Title: Exercise alters mRNA expression of telomere-repeat binding factor 1 in skeletal muscle via p38 MAPK

    doi: 10.1152/japplphysiol.00200.2012

    Figure Lengend Snippet: Calcium ionophore stimulation results in p38 MAPK phosphorylation in C2C12 myotubes. To determine whether calcium ionophore treatment resulted in activation/phosphorylation of p38 MAPK, myotubes were treated for 3 h with 1 μM A23187 or control, and protein was isolated 30 min and 24 h following treatment. Calcium ionophore treatment resulted in significant phosphorylation of p38 MAPK compared with BL. Densitometric analysis ( top ) and representative immunoblot images ( bottom ) are shown. Values are means ± SE. Phosphorylated-to-total protein content ratios were derived and then expressed relative to the BL group for comparisons. GAPDH was a loading reference. BL, vehicle treated; A23187, treated for 3 h, and protein isolated 30 min posttreatment; 24H post, myotubes treated for 3 h, protein isolated 24 h posttreatment. All experiments were performed in duplicate on at least 3 separate occasions, in serial passage. *A23187 significantly different than BL ( P

    Article Snippet: For total protein isoforms and GAPDH analyses, membranes from phosphorylated immunoblots were stripped and reprobed with the following antibodies for total proteins: total p38 MAPK (Cell Signaling 9212, 1:500), total ERK1/2 (Cell Signaling 9102, 1:1,000), and GAPDH (Cell Signaling, Danvers, MA, 14C10 rabbit MAb no. 2118, 1: 1,000).

    Techniques: Activation Assay, Isolation, Derivative Assay

    p38 MAPK is activated following acute treadmill exercise in the plantaris. We measured the phosphorylation status of the MAPKs ERK1/2 ( A ) and p38 ( B ) following acute treadmill exercise. ERK1/2 was not different among any groups (ERK1 p44, P = 0.6 and ERK2 p42, P = 0.8). p38 MAPK had significantly increased phosphorylation at TP1 compared with BL ( P = 0.03), TP1 and TP2 were similar ( P = 0.2), as were BL and TP2 ( P = 0.3). Densitometric analysis and representative immunoblot images ( right ) are shown. Values are means ± SE. Phosphorylated (p)-to-total (t) protein content ratios were derived and then expressed relative to the BL group for comparisons. GAPDH was a loading reference. BL, n = 6; TP1, n = 8; TP2, n = 8. *TP1 significantly different than BL ( P

    Journal: Journal of Applied Physiology

    Article Title: Exercise alters mRNA expression of telomere-repeat binding factor 1 in skeletal muscle via p38 MAPK

    doi: 10.1152/japplphysiol.00200.2012

    Figure Lengend Snippet: p38 MAPK is activated following acute treadmill exercise in the plantaris. We measured the phosphorylation status of the MAPKs ERK1/2 ( A ) and p38 ( B ) following acute treadmill exercise. ERK1/2 was not different among any groups (ERK1 p44, P = 0.6 and ERK2 p42, P = 0.8). p38 MAPK had significantly increased phosphorylation at TP1 compared with BL ( P = 0.03), TP1 and TP2 were similar ( P = 0.2), as were BL and TP2 ( P = 0.3). Densitometric analysis and representative immunoblot images ( right ) are shown. Values are means ± SE. Phosphorylated (p)-to-total (t) protein content ratios were derived and then expressed relative to the BL group for comparisons. GAPDH was a loading reference. BL, n = 6; TP1, n = 8; TP2, n = 8. *TP1 significantly different than BL ( P

    Article Snippet: For total protein isoforms and GAPDH analyses, membranes from phosphorylated immunoblots were stripped and reprobed with the following antibodies for total proteins: total p38 MAPK (Cell Signaling 9212, 1:500), total ERK1/2 (Cell Signaling 9102, 1:1,000), and GAPDH (Cell Signaling, Danvers, MA, 14C10 rabbit MAb no. 2118, 1: 1,000).

    Techniques: Derivative Assay

    p38 MAPK is not activated following acute treadmill exercise in the TA. A : ERK1 (p44) phosphorylation was not significantly altered in the TA due to treadmill exercise ( P = 0.4). ERK2 (p42) phosphorylation was greater at TP1 compared with TP2 ( P = 0.04) and tended to be greater than BL ( P = 0.08), but BL and TP2 were similar ( P = 0.8). #Significantly different than TP2 ( P

    Journal: Journal of Applied Physiology

    Article Title: Exercise alters mRNA expression of telomere-repeat binding factor 1 in skeletal muscle via p38 MAPK

    doi: 10.1152/japplphysiol.00200.2012

    Figure Lengend Snippet: p38 MAPK is not activated following acute treadmill exercise in the TA. A : ERK1 (p44) phosphorylation was not significantly altered in the TA due to treadmill exercise ( P = 0.4). ERK2 (p42) phosphorylation was greater at TP1 compared with TP2 ( P = 0.04) and tended to be greater than BL ( P = 0.08), but BL and TP2 were similar ( P = 0.8). #Significantly different than TP2 ( P

    Article Snippet: For total protein isoforms and GAPDH analyses, membranes from phosphorylated immunoblots were stripped and reprobed with the following antibodies for total proteins: total p38 MAPK (Cell Signaling 9212, 1:500), total ERK1/2 (Cell Signaling 9102, 1:1,000), and GAPDH (Cell Signaling, Danvers, MA, 14C10 rabbit MAb no. 2118, 1: 1,000).

    Techniques:

    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.

    Journal: The Journal of Experimental Medicine

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

    doi: 10.1084/jem.20091776

    Figure Lengend 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.

    Article Snippet: Activation of STAT-1, Akt, ERK, JNK, p38, or IKK-α was analyzed by immunoblot with the phosphospecific antibody for STAT-1 (Tyr701; 9171; Cell Signaling Technology), Akt (Ser473; 9271; Cell Signaling Technology), ERK (Tyr204; sc-7383; Santa Cruz Biotechnology, Inc.), JNK (Thr183/Tyr185; 9255; Cell Signaling Technology), p38 (Thr180/Tyr182; 9211; Cell Signaling Technology), or IKK-α (Ser176/180; 2697; Cell Signaling Technology), respectively.

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

    OSM-induced Activation of the MAPK Pathways in Primary Astrocytes A–C. Primary astrocytes were treated with OSM (5 ng/ml) for 0–24 h. Protein lysates were prepared, separated by SDS-PAGE, and subjected to immunoblot analysis with antibodies against (A) phosphorylated p38 Thr180/Tyr182 , (B) SAPK/JNK Thr183/Tyr182 or (C) p44/42 ERK Thr202/Tyr204 . Membranes were stripped and reprobed for total p38, SAPK/JNK, and p44/42 ERK, respectively, and actin as a loading control. Representative of three experiments.

    Journal: Glia

    Article Title: Molecular Basis of Oncostatin M-induced SOCS-3 Expression in Astrocytes

    doi: 10.1002/glia.20694

    Figure Lengend Snippet: OSM-induced Activation of the MAPK Pathways in Primary Astrocytes A–C. Primary astrocytes were treated with OSM (5 ng/ml) for 0–24 h. Protein lysates were prepared, separated by SDS-PAGE, and subjected to immunoblot analysis with antibodies against (A) phosphorylated p38 Thr180/Tyr182 , (B) SAPK/JNK Thr183/Tyr182 or (C) p44/42 ERK Thr202/Tyr204 . Membranes were stripped and reprobed for total p38, SAPK/JNK, and p44/42 ERK, respectively, and actin as a loading control. Representative of three experiments.

    Article Snippet: Abs against phospho-Jak-1Tyr1022/Tyr1023 , phospho-STAT-1αTyr701 , phospho-STAT-3Tyr705 , phospho-p44/42Thr202/Tyr204 , phospho-SAPK/JNKThr183/Tyr185 , phospho-p38 Thr180/Tyr182 , and p44/42, were obtained from Cell Signaling Technology (Danvers, MA).

    Techniques: Activation Assay, SDS Page

    Dasatinib inhibits PDGFR-β, c-Kit and c-Src phosphorylation in mesenchymal and osteoblast-like cell lines. (A) Mesenchymal (hMSC-TERT) and osteoblast-like (MG-63) cell lines were pretreated with different concentrations of dasatinib for 6 hours and then exposed to PDGF-BB or SCF for 20 minutes before protein lysates were generated. Immunoblotting with specific antibodies against total and phosphorylated PDGFR-β, c-Kit and c-Src were performed. (B) Modulation of downstream signaling after dasatinib treatment. Similarly to experimental conditions in (A), the hMSC-TERT and the MG-63 cell lines were pretreated with 50 nM dasatinib for 6 hours, stimulated with PDGF-BB for 20 minutes and then cell harvested for protein isolation. Immunoblotting is shown for total and phosphorylated forms of PDGFR-β, c-Src, Erk 1/2, Akt and p38 mitogen activated protein kinase (MAPK).

    Journal: PLoS ONE

    Article Title: Dasatinib as a Bone-Modifying Agent: Anabolic and Anti-Resorptive Effects

    doi: 10.1371/journal.pone.0034914

    Figure Lengend Snippet: Dasatinib inhibits PDGFR-β, c-Kit and c-Src phosphorylation in mesenchymal and osteoblast-like cell lines. (A) Mesenchymal (hMSC-TERT) and osteoblast-like (MG-63) cell lines were pretreated with different concentrations of dasatinib for 6 hours and then exposed to PDGF-BB or SCF for 20 minutes before protein lysates were generated. Immunoblotting with specific antibodies against total and phosphorylated PDGFR-β, c-Kit and c-Src were performed. (B) Modulation of downstream signaling after dasatinib treatment. Similarly to experimental conditions in (A), the hMSC-TERT and the MG-63 cell lines were pretreated with 50 nM dasatinib for 6 hours, stimulated with PDGF-BB for 20 minutes and then cell harvested for protein isolation. Immunoblotting is shown for total and phosphorylated forms of PDGFR-β, c-Src, Erk 1/2, Akt and p38 mitogen activated protein kinase (MAPK).

    Article Snippet: Primary antibodies for immunoblotting, immunohistochemical and flow cytometry analyses were directed against: PDGFR-β, phospho-PDGFR-β (Tyr857), Erk1/2, phospho-Erk1/2 (Thr202/Tyr204), NFATc1, histone H1 and cathepsin K, purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); phospho-c-Fms (Tyr723), phospho-c-Kit (Tyr719), c-Src, phospho-Src (Tyr416), p38 MAPK, phospho-p38 MAPK (Thr180/Tyr182), Akt, phospho-Akt (Ser473), phospho-β-catenin (Thr41/Ser45), PU.1 and c-Fos, from Cell Signaling Technology (Danvers, MA, USA); CD51/61 and CD191, from R & D Systems (Minneapolis, MN, USA); c-Kit and nucleoporin p62, from BD Biosciences (Bedford, MA, USA); α-tubulin, from Calbiochem (Darmstadt, Germany); dephospho-β-catenin (aa 35–50), from Enzo Life Sciences (Plymouth Meeting, PA, USA), and T-cell factor 4 (Tcf4), from Upstate (Millipore, Billerica, MA, USA).

    Techniques: Generated, Isolation

    Proposed mechanism by which OA-NO 2 reduces fibrosis in the atrium of AngII-treated mice. OA-NO 2 decreases fibroblast transdifferentiation through inhibition of Smad-2 activation, as indicated by a reduction of α-SMA expression. Additionally, OA-NO 2 reduces atrial oxidative stress due to attenuation of superoxide production from macrophages and fibroblasts by inhibition of NOX2 and p67 phox expression via p38 MAPK deactivation.

    Journal: Cardiovascular Research

    Article Title: Nitrated fatty acids suppress angiotensin II-mediated fibrotic remodelling and atrial fibrillation

    doi: 10.1093/cvr/cvv254

    Figure Lengend Snippet: Proposed mechanism by which OA-NO 2 reduces fibrosis in the atrium of AngII-treated mice. OA-NO 2 decreases fibroblast transdifferentiation through inhibition of Smad-2 activation, as indicated by a reduction of α-SMA expression. Additionally, OA-NO 2 reduces atrial oxidative stress due to attenuation of superoxide production from macrophages and fibroblasts by inhibition of NOX2 and p67 phox expression via p38 MAPK deactivation.

    Article Snippet: The protein was then transferred to polyvinylidene difluoride (PVDF) membranes (Immobilon-P, Millipore, Billerica, MA, USA) and incubated with antibodies against α-smooth muscle actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p38 MAPK, phospho-p38 MAPK (Thr180/Tyr182), Smad2, and phospho-Smad2 (Ser 465/467) (all purchased from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Mouse Assay, Inhibition, Activation Assay, Expressing

    OA-NO 2 inhibits atrial generation of reactive inflammatory mediators. ( A and B ) DHE stainings of atrial sections reveal a significantly reduced ROS formation in AngII(+)/OA-NO 2 (+) mice ( n = 8) compared with AngII(+)/OA-NO 2 (−) animals ( n = 7), with ROS levels in AngII(+)/OA-NO 2 (+) equalling those of control animals ( n = 8). Scale bar indicates 750 µm. ( C ) TGF-β-induced superoxide production measured by cytochrome c reduction assay in 3T3 fibroblasts is significantly blunted upon OA-NO 2 treatment ( n = 5). ( D ) LPS-induced production of superoxide in murine peritoneal RAW 264.7 macrophages measured by cytochrome c reduction assay is inhibited by OA-NO 2 in a dose-dependent manner ( n = 6). ( E ) Oxidative burst in murine peritoneal RAW 264.7 macrophages induced by phorbol-12-myristate-13-acetate (PMA) is also reduced by application of OA-NO 2 ( n = 4). ( F and G ) The LPS-induced expression of NOX2 as well as of its cytosolic regulatory protein p67phox is significantly attenuated by OA-NO 2 treatment in a dose-dependent manner ( n = 5). ( H and I ) NOX2 stainings of atrial sections reveal a significantly reduced NOX2 expression in AngII(+)/OA-NO 2 (+) mice ( n = 8) compared with AngII(+)/OA-NO 2 (−) animals ( n = 7), with ROS levels in AngII(+)/OA-NO 2 (+) equalling those of control animals ( n = 8). Scale bar indicates 50 µm. ( J ) OA-NO 2 decreases phosphorylation of p38 MAPK. Pooled data from four independent experiments are shown. Data are expressed as mean ± SEM. * P

    Journal: Cardiovascular Research

    Article Title: Nitrated fatty acids suppress angiotensin II-mediated fibrotic remodelling and atrial fibrillation

    doi: 10.1093/cvr/cvv254

    Figure Lengend Snippet: OA-NO 2 inhibits atrial generation of reactive inflammatory mediators. ( A and B ) DHE stainings of atrial sections reveal a significantly reduced ROS formation in AngII(+)/OA-NO 2 (+) mice ( n = 8) compared with AngII(+)/OA-NO 2 (−) animals ( n = 7), with ROS levels in AngII(+)/OA-NO 2 (+) equalling those of control animals ( n = 8). Scale bar indicates 750 µm. ( C ) TGF-β-induced superoxide production measured by cytochrome c reduction assay in 3T3 fibroblasts is significantly blunted upon OA-NO 2 treatment ( n = 5). ( D ) LPS-induced production of superoxide in murine peritoneal RAW 264.7 macrophages measured by cytochrome c reduction assay is inhibited by OA-NO 2 in a dose-dependent manner ( n = 6). ( E ) Oxidative burst in murine peritoneal RAW 264.7 macrophages induced by phorbol-12-myristate-13-acetate (PMA) is also reduced by application of OA-NO 2 ( n = 4). ( F and G ) The LPS-induced expression of NOX2 as well as of its cytosolic regulatory protein p67phox is significantly attenuated by OA-NO 2 treatment in a dose-dependent manner ( n = 5). ( H and I ) NOX2 stainings of atrial sections reveal a significantly reduced NOX2 expression in AngII(+)/OA-NO 2 (+) mice ( n = 8) compared with AngII(+)/OA-NO 2 (−) animals ( n = 7), with ROS levels in AngII(+)/OA-NO 2 (+) equalling those of control animals ( n = 8). Scale bar indicates 50 µm. ( J ) OA-NO 2 decreases phosphorylation of p38 MAPK. Pooled data from four independent experiments are shown. Data are expressed as mean ± SEM. * P

    Article Snippet: The protein was then transferred to polyvinylidene difluoride (PVDF) membranes (Immobilon-P, Millipore, Billerica, MA, USA) and incubated with antibodies against α-smooth muscle actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p38 MAPK, phospho-p38 MAPK (Thr180/Tyr182), Smad2, and phospho-Smad2 (Ser 465/467) (all purchased from Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Mouse Assay, Expressing

    DHX9 is not required for TLR2-stimulated NF-κB or p38 MAPK activation. THP-1 cells expressing doxycycline-inducible DHX9-specific shRNA (G2) or NSC shRNA, were incubated with 1 μg/ml of doxycycline for 72 h, and then stimulated with 2

    Journal: The Journal of Biological Chemistry

    Article Title: Poxviral protein E3–altered cytokine production reveals that DExD/H-box helicase 9 controls Toll-like receptor–stimulated immune responses

    doi: 10.1074/jbc.RA118.005089

    Figure Lengend Snippet: DHX9 is not required for TLR2-stimulated NF-κB or p38 MAPK activation. THP-1 cells expressing doxycycline-inducible DHX9-specific shRNA (G2) or NSC shRNA, were incubated with 1 μg/ml of doxycycline for 72 h, and then stimulated with 2

    Article Snippet: Antibodies used were anti-HA (mouse, Covance), anti-β-actin (mouse, Sigma), anti-E3 (a gift from B. Jacobs ASU Biodesign Institute, Arizona State University), anti-DHX9 (rabbit, Novus Biologicals), anti-tubulin (mouse, Millipore), anti-IκBα (mouse, gift from R. T. Hay, University of Dundee), anti-phospho-Ser536 -p65 (rabbit, Cell Signaling), anti-p65 (mouse, Santa Cruz), anti-phospho-p38 MAPK (rabbit, Cell Signaling), and anti-p38 MAPK (rabbit, Cell Signaling).

    Techniques: Activation Assay, Expressing, shRNA, Incubation

    Activation of MAPKs on human breast carcinoma MCF-7 cells treated with PMA. Cells were incubated in the presence of PMA (50 ng/ml) during the indicated times, and the activation status of ERK1/2, p38, and JNK determined using phospho-specific anti-ERK1/2

    Journal: The Journal of Biological Chemistry

    Article Title: Differential Up-regulation of MAP Kinase Phosphatases MKP3/DUSP6 and DUSP5 by Ets2 and c-Jun Converge in the Control of the Growth Arrest Versus Proliferation Response of MCF-7 Breast Cancer Cells to Phorbol Ester *

    doi: 10.1074/jbc.M110.121830

    Figure Lengend Snippet: Activation of MAPKs on human breast carcinoma MCF-7 cells treated with PMA. Cells were incubated in the presence of PMA (50 ng/ml) during the indicated times, and the activation status of ERK1/2, p38, and JNK determined using phospho-specific anti-ERK1/2

    Article Snippet: Other polyclonal antibodies used were: anti-DUSP5 (provided by Steve Keyse ( )), anti-MKP3 to detect endogenous MKP3 ( and ) (provided by Johan Lennartsson , anti-pp38, anti-pAKT, anti-AKT (Cell Signaling), anti-Ets2, anti-c-Jun, anti-ERK1/2, anti-p38, and anti-JNK (all from Santa Cruz), and anti-Ets2 (Thr(P)-72) (Invitrogen).

    Techniques: Activation Assay, Incubation

    Survival pathway analysis. TUDCA treatment upregulated ERK1/2 following ischemia-reperfusion injury to the kidney in rats (top panel); however, there was no statistical significant difefrence as compared to the vehicle-treated rats (top panel). There was no difference in JNK and p38 proteins between the TUDCA and vehicle groups. Densitometry analysis of ERK1/2 in each rat in the TUDCA group were compared with those in the vehicle group (lower panel); the difference was not significant ( p = 0.29). Results are expressed as mean ± standard deviation of a least 3 different animals in each group.

    Journal: PLoS ONE

    Article Title: Prevention of Acute Kidney Injury by Tauroursodeoxycholic Acid in Rat and Cell Culture Models

    doi: 10.1371/journal.pone.0048950

    Figure Lengend Snippet: Survival pathway analysis. TUDCA treatment upregulated ERK1/2 following ischemia-reperfusion injury to the kidney in rats (top panel); however, there was no statistical significant difefrence as compared to the vehicle-treated rats (top panel). There was no difference in JNK and p38 proteins between the TUDCA and vehicle groups. Densitometry analysis of ERK1/2 in each rat in the TUDCA group were compared with those in the vehicle group (lower panel); the difference was not significant ( p = 0.29). Results are expressed as mean ± standard deviation of a least 3 different animals in each group.

    Article Snippet: Anti-phospho-ERK1/2 antibody was purchased from New England BioLabs (Boston, MA); anti-phospho-p38 and anti-phospho-JNK from Santa Cruz Biotechnology Inc. (Santa Cruz, CA); anti-caspase-8 and anti-caspase-12 from BioVision Inc. (Mountainview, CA); anti-caspase-9 antibody from Enzo Life Sciences (Plymouth Meeting, PA); and anti-mouse β-actin from Calbiochem (Spring Valley,CA).

    Techniques: Standard Deviation

    The p38 cascade is up-regulated in patients with relapsing–remitting multiple sclerosis (RR-MS): (a) Graph represents the median fluorescence intensity (MFI) of p38 total protein (left) and phosphorylated protein (right) in peripheral blood mononuclear

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: The p38 cascade is up-regulated in patients with relapsing–remitting multiple sclerosis (RR-MS): (a) Graph represents the median fluorescence intensity (MFI) of p38 total protein (left) and phosphorylated protein (right) in peripheral blood mononuclear

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Mass Spectrometry, Fluorescence

    p38 activation is required for interleukin-17 (IL-17) release in patients with relapsing–remitting multiple sclerosis (RR-MS): FACS-sorted CD4 + CD45RA + CD27 + naive T cells isolated from patients with RR-MS were polarized into T

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: p38 activation is required for interleukin-17 (IL-17) release in patients with relapsing–remitting multiple sclerosis (RR-MS): FACS-sorted CD4 + CD45RA + CD27 + naive T cells isolated from patients with RR-MS were polarized into T

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Activation Assay, Mass Spectrometry, FACS, Isolation

    p38 activation is essential for T helper type 17 (Th17) generation from human CD4 + naive T cells in vitro : (a) FACS-sorted CD4 + CD45RA + CD27 + naive T cells were stimulated with anti-CD3 and anti-CD28 in the presence (lower panel)

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: p38 activation is essential for T helper type 17 (Th17) generation from human CD4 + naive T cells in vitro : (a) FACS-sorted CD4 + CD45RA + CD27 + naive T cells were stimulated with anti-CD3 and anti-CD28 in the presence (lower panel)

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Activation Assay, In Vitro, FACS

    eIF-4E activation is required for T helper type 17 (Th17) generation and interleukin-17 (IL-17) release in human CD4 + naive cells: (a) FACS-sorted naive CD4 + T cells were stimulated for 20 min with PMA/ionomycin and then checked for p38 phosphorylation

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: eIF-4E activation is required for T helper type 17 (Th17) generation and interleukin-17 (IL-17) release in human CD4 + naive cells: (a) FACS-sorted naive CD4 + T cells were stimulated for 20 min with PMA/ionomycin and then checked for p38 phosphorylation

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Activation Assay, FACS

    p38 inhibition modulates interleukin-17 (IL-17) release from central memory T cells and T helper type 17 (Th17) cell clones: (a) FACS-sorted CD4 + CD45RA − CD27 + memory T cells were plated in the presence or absence of the Th17 cytokine

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: p38 inhibition modulates interleukin-17 (IL-17) release from central memory T cells and T helper type 17 (Th17) cell clones: (a) FACS-sorted CD4 + CD45RA − CD27 + memory T cells were plated in the presence or absence of the Th17 cytokine

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Inhibition, FACS

    p38 inhibition does not affect interferon- γ (IFN- γ ) and Foxp3 expression in culture and significantly increases interleukin-21 (IL-21) release: FACS-sorted CD4 + naive T lymphocytes were polarized into T helper type 17 (Th17) cells with

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: p38 inhibition does not affect interferon- γ (IFN- γ ) and Foxp3 expression in culture and significantly increases interleukin-21 (IL-21) release: FACS-sorted CD4 + naive T lymphocytes were polarized into T helper type 17 (Th17) cells with

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Inhibition, Expressing, FACS

    p38 activation modulates interleukin-17 (IL-17) release and RORC and IL-17 expression in vitro : (a) IL-17 levels in the supernatants from three distinct differentiation assays were assessed by ELISA. (b) FACS-sorted CD4 + CD45RA + CD27

    Journal: Immunology

    Article Title: The p38 mitogen-activated protein kinase cascade modulates T helper type 17 differentiation and functionality in multiple sclerosis

    doi: 10.1111/imm.12497

    Figure Lengend Snippet: p38 activation modulates interleukin-17 (IL-17) release and RORC and IL-17 expression in vitro : (a) IL-17 levels in the supernatants from three distinct differentiation assays were assessed by ELISA. (b) FACS-sorted CD4 + CD45RA + CD27

    Article Snippet: Cells were tested by Western blot analysis to detect the levels of rabbit anti-phospho-p38 (Thr180 and Tyr182; Cell Signaling), anti-phospho-eIF4E (Ser209, BioSource International, Camarillo, CA), and mouse anti β -tubulin (Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, In Vitro, Enzyme-linked Immunosorbent Assay, FACS

    PKC mediated activation of MAPK and AKT pathways by KA stimulation. (A) Western blot analysis of time-dependent activation of JAK1, STAT1, AKT, MAPK ERK1/2 and p38 after adding 100μM KA to 8 div hippocampal neurons for 30min. (B) Pretreatment of 8 div hippocampal neurons with PKC inhibitor (Staurosporine, 0.1μM) compromised the KA-induced activation of AKT, MAPK ERK1/2 and p38, but had no effect on the activation of JAK1 and STAT1. (C) Pretreatment of 8 div hippocampal neurons with PKA inhibitor (H89, 40μM) had no effect on the activation of JAK1, STAT1, AKT, MAPK ERK1/2 and p38 induced by KA. (D) Pretreatment of primary hippocampal neurons with MAPK-p38 inhibitor (Skepinone-L, 20μM) resulted in inhibition of KA-induced expression of MHC-I. (E) Pretreatment of primary hippocampal neurons with MAPK-ERK inhibitor (U0126, 10μM) blocked KA-induced expression of MHC-I. (F) Pretreatment of primary hippocampal neurons with MAPK-p38 inhibitor (Skepinone-L, 20μM) blocked KA-induced expression of p-p65, p-CREB and IRF-1. (G) Pretreatment of primary hippocampal neurons with MAPK-ERK inhibitor (U0126, 10μM) resulted in inhibition of KA-induced expression of IRF-1 and activation of NF-κB p65, but had no effect on the activation of CREB.

    Journal: PLoS ONE

    Article Title: Neuronal MHC Class I Expression Is Regulated by Activity Driven Calcium Signaling

    doi: 10.1371/journal.pone.0135223

    Figure Lengend Snippet: PKC mediated activation of MAPK and AKT pathways by KA stimulation. (A) Western blot analysis of time-dependent activation of JAK1, STAT1, AKT, MAPK ERK1/2 and p38 after adding 100μM KA to 8 div hippocampal neurons for 30min. (B) Pretreatment of 8 div hippocampal neurons with PKC inhibitor (Staurosporine, 0.1μM) compromised the KA-induced activation of AKT, MAPK ERK1/2 and p38, but had no effect on the activation of JAK1 and STAT1. (C) Pretreatment of 8 div hippocampal neurons with PKA inhibitor (H89, 40μM) had no effect on the activation of JAK1, STAT1, AKT, MAPK ERK1/2 and p38 induced by KA. (D) Pretreatment of primary hippocampal neurons with MAPK-p38 inhibitor (Skepinone-L, 20μM) resulted in inhibition of KA-induced expression of MHC-I. (E) Pretreatment of primary hippocampal neurons with MAPK-ERK inhibitor (U0126, 10μM) blocked KA-induced expression of MHC-I. (F) Pretreatment of primary hippocampal neurons with MAPK-p38 inhibitor (Skepinone-L, 20μM) blocked KA-induced expression of p-p65, p-CREB and IRF-1. (G) Pretreatment of primary hippocampal neurons with MAPK-ERK inhibitor (U0126, 10μM) resulted in inhibition of KA-induced expression of IRF-1 and activation of NF-κB p65, but had no effect on the activation of CREB.

    Article Snippet: The primary antibodies and concentrations used were as follows: mouse anti-MHC-I (ox18, 1:1000; Abcam), rabbit anti-IRF-1 antibody (1:500; Santa Cruz, Dallas, TX, USA), rabbit anti-CREB and p-CREB (ser133) antibody (1:1000; Cell Signaling), rabbit anti-p65 and p-p65 (Ser536) antibody (1:1000; Cell Signaling), rabbit anti-JAK1 and p-JAK1 (Y1022) antibody (1:1000, Bioworld, St. Louis Park, MN, USA), rabbit anti-STAT1 and p-STAT1 (Tyr701) antibody (1:1000; Cell Signaling), rabbit anti-p38 and p-p38 antibody (1:1000; Cell Signaling), rabbit anti-ERK1/2 and p-ERK1/2 (Thr202/Tyr204) antibody (1:1000, Cell Signaling), rabbit anti-AKT and p-AKT (Ser473) antibody (1:1000; Cell Signaling), rabbit anti-synaptophysin antibody (1:1000, Santa Cruz), mouse anti-PSD-95 antibody (1:500, Thermo Fisher Scientific, Rockford, IL, USA), rabbit anti-c-fos antibody (1:500, Santa Cruz).

    Techniques: Activation Assay, Western Blot, Inhibition, Expressing

    APIM-peptide increases the effects of docetaxel on apoptosis and cellular signaling ( A ) Cell cycle analysis and fraction of apoptotic (A) vs necrotic (N) cells (values from contour plots are given). ( B ) Relative expression of phosphorylated Akt, ERK1, ERK2, S6K, and p38, cleaved-PARP, and caspase 3. For all experiments, PC3 and Du145 cells were treated with APIM-peptide (14 μg/mL, green), docetaxel (2.5 ng/mL: PC3; 1.3 ng/mL: Du145; 5 ng/mL, blue) and the combination of these (black) for 24 hours prior to the analysis. The protein levels (B) were adjusted for loading differences (β-tubulin) and normalized against untreated cells, and additionally for total protein levels for the phosphorylated proteins. Data are from three biological replicas (different symbols represents extracts acquired on different days). Statistically significant differences ( * p

    Journal: Oncotarget

    Article Title: APIM-peptide targeting PCNA improves the efficacy of docetaxel treatment in the TRAMP mouse model of prostate cancer

    doi: 10.18632/oncotarget.24357

    Figure Lengend Snippet: APIM-peptide increases the effects of docetaxel on apoptosis and cellular signaling ( A ) Cell cycle analysis and fraction of apoptotic (A) vs necrotic (N) cells (values from contour plots are given). ( B ) Relative expression of phosphorylated Akt, ERK1, ERK2, S6K, and p38, cleaved-PARP, and caspase 3. For all experiments, PC3 and Du145 cells were treated with APIM-peptide (14 μg/mL, green), docetaxel (2.5 ng/mL: PC3; 1.3 ng/mL: Du145; 5 ng/mL, blue) and the combination of these (black) for 24 hours prior to the analysis. The protein levels (B) were adjusted for loading differences (β-tubulin) and normalized against untreated cells, and additionally for total protein levels for the phosphorylated proteins. Data are from three biological replicas (different symbols represents extracts acquired on different days). Statistically significant differences ( * p

    Article Snippet: The membranes were blocked in blocking buffer (5% dry milk in PBS) before incubation with primary antibodies against Akt (Cell Signaling Technology, #4691, MA, USA), Phospo-Akt (P-Akt, Ser473, Cell Signaling Technology, #4060, MA, USA), ERK1/2 (ERK, Santa Cruz Biotechnology, sc-93-G, TX, USA), Phospho-ERK1/2 (P-ERK, Thr202/Tyr204, #4370, Cell Signaling Technology, MA, USA), p70 S6 Kinase (S6K, Cell Signaling Technology, #2708, MA, USA), Phospho-S6K (P-S6K, Thr389, Cell Signaling Technology, #9206, MA, USA), p38 (Abcam, ab31828, UK), cleaved PARP (Abcam, ab4830, UK), Caspase 3 (Cell Signaling Technology, #9662, MA, USA), β-tubulin (Abcam, ab6046, UK) and β-actin (Abcam, ab8226, UK).

    Techniques: Cell Cycle Assay, Expressing

    Activated p38 downregulates the stem cell properties of NSCLC cells ( A ) Flow cytometry showing the percentage of the side population detected by Hoechst33342 staining in A549 and H460 cells transduced with vector control (BP), MKK3E or MKK6E and H460 and H1299 cells transduced with vector control (BH), MKK3A or MKK6A. Bar graphs show quantification of the percentage of side population. ( B ) Sphere formation assay of A549 and H460 cells transduced with vector control (BP), MKK3E or MKK6E and H460 and H1299 cells transduced with vector control (BH), MKK3A or MKK6A. Scar bar, 100 μm. Bar graphs show quantifications of the number and diameter of the spheres, respectively. * indicates P

    Journal: Oncotarget

    Article Title: Inactivation of p38 MAPK contributes to stem cell-like properties of non-small cell lung cancer

    doi: 10.18632/oncotarget.15804

    Figure Lengend Snippet: Activated p38 downregulates the stem cell properties of NSCLC cells ( A ) Flow cytometry showing the percentage of the side population detected by Hoechst33342 staining in A549 and H460 cells transduced with vector control (BP), MKK3E or MKK6E and H460 and H1299 cells transduced with vector control (BH), MKK3A or MKK6A. Bar graphs show quantification of the percentage of side population. ( B ) Sphere formation assay of A549 and H460 cells transduced with vector control (BP), MKK3E or MKK6E and H460 and H1299 cells transduced with vector control (BH), MKK3A or MKK6A. Scar bar, 100 μm. Bar graphs show quantifications of the number and diameter of the spheres, respectively. * indicates P

    Article Snippet: The antibodies used in this study including anti-β-actin (Santa cruz; sc-47778), anti-p38 (abcam; ab32142), anti-phopho-p38 MAPK(Cell signaling Technology; #9215), anti-SOX2 (Santa cruz; sc-20088), anti-Oct4 (abcam; ab19857), anti-HA (Cell signaling Technology, #3724), anti-c-Myc (Cell signaling Technology; #13987), anti-Nanog (abcam; ab80892), anti-Klf4 (Cell signaling Technology, #4038), anti-ERK (Beijing Zhongshan Golden Bridge Biotechnology Co Ltd, ZS94), anti-Ub (Santa cruz, sc-8017), anti-phospho-ATF2 (Cell signaling Technology, #9221).

    Techniques: Flow Cytometry, Cytometry, Staining, Transduction, Plasmid Preparation, Tube Formation Assay

    Activated p38γ and p38δ suppress the stem cell properties of NSCLC cells ( A ) Western blot analysis of the stemness markers in A549 and H460 cells transduced with vector control (BP), p38aWT, p38αD176A, p38bWT, p38βD176A, p38gWT, p38γD179A, p38dWT, or p38δF324S, and in H460 and H1299 cells transduced with shRNA for GFP or indicated p38 isoforms. ( B ) Quantification of percentage of the side population in A549 and H460 cells transduced with vector control (BP), p38aWT, p38αD176A, p38bWT, p38βD176A, p38gWT, p38γD179A, p38dWT, or p38δF324S, and in H460 and H1299 cells transduced with shRNA for GFP or indicated p38 isoforms, as determined by flow cytometry. ( C ) Limit dilution xenograft tumor formation assay, showing number of tumors arising after subcutaneous injection of indicated numbers of A549 cells transduced with vector control (BP), p38γD179A or p38δF324S into 8 injection sites in Nod-scid mice. ( D , E ) The growth curves of xenograft tumors formed by 1 × 10 6 (D) or 5 × 10 5 (E) of A549 cells transduced with vector control (BP), p38γD179A or p38δF324S. Tumor volumes = length × width 2 /2. * indicates P

    Journal: Oncotarget

    Article Title: Inactivation of p38 MAPK contributes to stem cell-like properties of non-small cell lung cancer

    doi: 10.18632/oncotarget.15804

    Figure Lengend Snippet: Activated p38γ and p38δ suppress the stem cell properties of NSCLC cells ( A ) Western blot analysis of the stemness markers in A549 and H460 cells transduced with vector control (BP), p38aWT, p38αD176A, p38bWT, p38βD176A, p38gWT, p38γD179A, p38dWT, or p38δF324S, and in H460 and H1299 cells transduced with shRNA for GFP or indicated p38 isoforms. ( B ) Quantification of percentage of the side population in A549 and H460 cells transduced with vector control (BP), p38aWT, p38αD176A, p38bWT, p38βD176A, p38gWT, p38γD179A, p38dWT, or p38δF324S, and in H460 and H1299 cells transduced with shRNA for GFP or indicated p38 isoforms, as determined by flow cytometry. ( C ) Limit dilution xenograft tumor formation assay, showing number of tumors arising after subcutaneous injection of indicated numbers of A549 cells transduced with vector control (BP), p38γD179A or p38δF324S into 8 injection sites in Nod-scid mice. ( D , E ) The growth curves of xenograft tumors formed by 1 × 10 6 (D) or 5 × 10 5 (E) of A549 cells transduced with vector control (BP), p38γD179A or p38δF324S. Tumor volumes = length × width 2 /2. * indicates P

    Article Snippet: The antibodies used in this study including anti-β-actin (Santa cruz; sc-47778), anti-p38 (abcam; ab32142), anti-phopho-p38 MAPK(Cell signaling Technology; #9215), anti-SOX2 (Santa cruz; sc-20088), anti-Oct4 (abcam; ab19857), anti-HA (Cell signaling Technology, #3724), anti-c-Myc (Cell signaling Technology; #13987), anti-Nanog (abcam; ab80892), anti-Klf4 (Cell signaling Technology, #4038), anti-ERK (Beijing Zhongshan Golden Bridge Biotechnology Co Ltd, ZS94), anti-Ub (Santa cruz, sc-8017), anti-phospho-ATF2 (Cell signaling Technology, #9221).

    Techniques: Western Blot, Transduction, Plasmid Preparation, shRNA, Flow Cytometry, Cytometry, Tube Formation Assay, Injection, Mouse Assay

    Activated p38 MAPK reduces protein stability of the stemness proteins and promotes ubiquitylation and proteasome-mediated degradation of SOX2 ( A ) Relative mRNA levels of the stemness proteins in A549 cells transduced with vector control (BP), MKK3E or MKK6E and H1299 cells transduced with vector control (BH) or MKK6A, as determined by quantitative real time PCR analysis. ns indicates no significant difference with P > 0.05, * indicates significant difference with P

    Journal: Oncotarget

    Article Title: Inactivation of p38 MAPK contributes to stem cell-like properties of non-small cell lung cancer

    doi: 10.18632/oncotarget.15804

    Figure Lengend Snippet: Activated p38 MAPK reduces protein stability of the stemness proteins and promotes ubiquitylation and proteasome-mediated degradation of SOX2 ( A ) Relative mRNA levels of the stemness proteins in A549 cells transduced with vector control (BP), MKK3E or MKK6E and H1299 cells transduced with vector control (BH) or MKK6A, as determined by quantitative real time PCR analysis. ns indicates no significant difference with P > 0.05, * indicates significant difference with P

    Article Snippet: The antibodies used in this study including anti-β-actin (Santa cruz; sc-47778), anti-p38 (abcam; ab32142), anti-phopho-p38 MAPK(Cell signaling Technology; #9215), anti-SOX2 (Santa cruz; sc-20088), anti-Oct4 (abcam; ab19857), anti-HA (Cell signaling Technology, #3724), anti-c-Myc (Cell signaling Technology; #13987), anti-Nanog (abcam; ab80892), anti-Klf4 (Cell signaling Technology, #4038), anti-ERK (Beijing Zhongshan Golden Bridge Biotechnology Co Ltd, ZS94), anti-Ub (Santa cruz, sc-8017), anti-phospho-ATF2 (Cell signaling Technology, #9221).

    Techniques: Transduction, Plasmid Preparation, Real-time Polymerase Chain Reaction

    Activated p38 suppresses the stem cell-like properties of NSCLC cells through MK2-dependent phosphorylation of Hsp27 at Ser78 and Ser82 ( A ) Western blot analysis of Hsp27 phosphorylated at Ser15, Ser78 and Ser82 in A549 cells transduced with vector control (BP), p38gWT, p38γD179A, p38dWT, p38δF324S, MKK3E or MKK6E, and H1299 cells transduced with vector control (BH), MKK6A, or shRNA for p38γ or p38δ. ( B ) Western blot analysis of Hsp27 phosphorylated at Ser15, Ser78 and Ser82 in A549 cells transduced with a scrambled shRNA (SC) or shRNA for PRAK or MK2, and p38γD179A or p38δF324S. ( C ) Western blot analysis of the stemness proteins in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphomimetic mutants of Hsp27 (Hsp27-S15D, Hsp27-DblD, Hsp27-TriD) (left panels), and in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphorylation-resistant mutants of Hsp27 (Hsp27-S15A, Hsp27-DblA, Hsp27-TriA) and MKK3E (right panels). ( D ) Flow cytometry analysis was performed to determine the percentage of the side population in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphomimetic mutants of Hsp27 (Hsp27-DblD, Hsp27-TriD), and in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphorylation-resistant mutants of Hsp27 (Hsp27-DblA, Hsp27-TriA) and MKK3E. ( E ) Quantification of the results in (D). *** indicates significant difference with P

    Journal: Oncotarget

    Article Title: Inactivation of p38 MAPK contributes to stem cell-like properties of non-small cell lung cancer

    doi: 10.18632/oncotarget.15804

    Figure Lengend Snippet: Activated p38 suppresses the stem cell-like properties of NSCLC cells through MK2-dependent phosphorylation of Hsp27 at Ser78 and Ser82 ( A ) Western blot analysis of Hsp27 phosphorylated at Ser15, Ser78 and Ser82 in A549 cells transduced with vector control (BP), p38gWT, p38γD179A, p38dWT, p38δF324S, MKK3E or MKK6E, and H1299 cells transduced with vector control (BH), MKK6A, or shRNA for p38γ or p38δ. ( B ) Western blot analysis of Hsp27 phosphorylated at Ser15, Ser78 and Ser82 in A549 cells transduced with a scrambled shRNA (SC) or shRNA for PRAK or MK2, and p38γD179A or p38δF324S. ( C ) Western blot analysis of the stemness proteins in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphomimetic mutants of Hsp27 (Hsp27-S15D, Hsp27-DblD, Hsp27-TriD) (left panels), and in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphorylation-resistant mutants of Hsp27 (Hsp27-S15A, Hsp27-DblA, Hsp27-TriA) and MKK3E (right panels). ( D ) Flow cytometry analysis was performed to determine the percentage of the side population in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphomimetic mutants of Hsp27 (Hsp27-DblD, Hsp27-TriD), and in A549 cells transduced with vector control (MCS), wild type Hsp27 (Hsp27-WT) or phosphorylation-resistant mutants of Hsp27 (Hsp27-DblA, Hsp27-TriA) and MKK3E. ( E ) Quantification of the results in (D). *** indicates significant difference with P

    Article Snippet: The antibodies used in this study including anti-β-actin (Santa cruz; sc-47778), anti-p38 (abcam; ab32142), anti-phopho-p38 MAPK(Cell signaling Technology; #9215), anti-SOX2 (Santa cruz; sc-20088), anti-Oct4 (abcam; ab19857), anti-HA (Cell signaling Technology, #3724), anti-c-Myc (Cell signaling Technology; #13987), anti-Nanog (abcam; ab80892), anti-Klf4 (Cell signaling Technology, #4038), anti-ERK (Beijing Zhongshan Golden Bridge Biotechnology Co Ltd, ZS94), anti-Ub (Santa cruz, sc-8017), anti-phospho-ATF2 (Cell signaling Technology, #9221).

    Techniques: Western Blot, Transduction, Plasmid Preparation, shRNA, Flow Cytometry, Cytometry

    Activated p38 negatively regulates the expression of stemness proteins in non-small cell lung cancer (NSCLC) ( A ) IHC staining of SOX2 and p-p38 in serial sections of a human lung cancer tissue array containing 153 intact NSCLC tissues and 31 NAT/normal lung tissues. The scatter diagram shows quantification of area of positive staining for SOX2 and p-p38 in normal and tumor tissues, which is calculated by multiplying staining area (scored as 1, 2, 3, and 4, 1: 0–25%, 2: 25%–50%, 3: 50%–75%, 4: 75%–100%, of positive tissue area) with staining intensity (scored as 1, 2, 3, and 4 based on color). *** indicates significant difference with P

    Journal: Oncotarget

    Article Title: Inactivation of p38 MAPK contributes to stem cell-like properties of non-small cell lung cancer

    doi: 10.18632/oncotarget.15804

    Figure Lengend Snippet: Activated p38 negatively regulates the expression of stemness proteins in non-small cell lung cancer (NSCLC) ( A ) IHC staining of SOX2 and p-p38 in serial sections of a human lung cancer tissue array containing 153 intact NSCLC tissues and 31 NAT/normal lung tissues. The scatter diagram shows quantification of area of positive staining for SOX2 and p-p38 in normal and tumor tissues, which is calculated by multiplying staining area (scored as 1, 2, 3, and 4, 1: 0–25%, 2: 25%–50%, 3: 50%–75%, 4: 75%–100%, of positive tissue area) with staining intensity (scored as 1, 2, 3, and 4 based on color). *** indicates significant difference with P

    Article Snippet: The antibodies used in this study including anti-β-actin (Santa cruz; sc-47778), anti-p38 (abcam; ab32142), anti-phopho-p38 MAPK(Cell signaling Technology; #9215), anti-SOX2 (Santa cruz; sc-20088), anti-Oct4 (abcam; ab19857), anti-HA (Cell signaling Technology, #3724), anti-c-Myc (Cell signaling Technology; #13987), anti-Nanog (abcam; ab80892), anti-Klf4 (Cell signaling Technology, #4038), anti-ERK (Beijing Zhongshan Golden Bridge Biotechnology Co Ltd, ZS94), anti-Ub (Santa cruz, sc-8017), anti-phospho-ATF2 (Cell signaling Technology, #9221).

    Techniques: Expressing, Immunohistochemistry, Staining