rabbit anti human jnk1  (Boster Bio)

Journal: Mediators of Inflammation

Article Title: Cross-Linking Mast Cell Specific Gangliosides Stimulates the Release of Newly Formed Lipid Mediators and Newly Synthesized Cytokines

doi: 10.1155/2016/9160540

Figure Lengend Snippet: Cross-linking GD1b derived gangliosides with mAbAA4 induced MAP kinase phosphorylation in mast cells. RBL-2H3 cells were either stimulated via Fc ε RI, where cells were sensitized with IgE anti-TNP and stimulated with DNP 48 -HSA (50 ng/mL) or incubated with mAbAA4 (1, 2.5, 5, and 10 μ g/mL) for 10 min. Total cell lysates were immunoblotted with antibodies against phospho-ERK1/2 (p-ERK1/2), ERK1/2, phospho-JNK1/2 (p-JNK1/2), JNK1/2, phospho-p38 (p-p38), p38, and α / β -tubulin (housekeeping protein) and the mean optical density of the bands was determined. Densitometry of the changes in expression and phosphorylation of proteins were corrected for α / β -tubulin. Data is expressed as the fold of nonstimulated (NS) cells. (a) Ratio of phosphorylated ERK1/2/total ERK1/2; (b) a representative blot from (a); (c) ratio of phosphorylated JNK1/2/total JNK1/2; (d) a representative blot from (c); (e) ratio of phosphorylated p38/total p38; (f) a representative blot from (e). Data is expressed as the mean ± SD of three independent experiments. ∗ P < 0.05 between experimental samples and the nonstimulated (NS) cells. # P < 0.05 between experimental samples and Fc ε RI stimulated cells.

Article Snippet: Rabbit polyclonal antibody anti-human phospho-cPLA 2 ; rabbit polyclonal antibody anti-human cPLA 2 ; rabbit mAb anti-human phospho-ERK1/2; rabbit mAb anti-rat ERK1/2; rabbit mAb anti-human phospho-JNK1/2; rabbit polyclonal antibody anti-human JNK1/2; rabbit mAb anti-human phospho-p38; rabbit polyclonal antibody anti-human anti-p38, and rabbit polyclonal antibody anti-human α / β -tubulin were purchased from Cell Signaling Technology Inc. (Danvers, MA).

Techniques: Derivative Assay, Incubation, Expressing

Journal: Cancer gene therapy

Article Title: c-Jun N-terminal kinase 2 suppresses pancreatic cancer growth and invasion and is opposed by c-Jun N-terminal kinase 1.

doi: 10.1038/s41417-020-00290-5

Figure Lengend Snippet: Fig. 6 EMT markers in dependency of JNK expression. A Immu- noblot analysis of N-Cadherin, E-Cadherin, Vimentin, and α-Tubulin in pancreatic cancer cell lines. β-Actin is used to confirm equal loading. B, C JNK2 downregulation leads to reduced Vimentin expression. MMP9 is upregulated in the KD of both isoforms, but predominantly in JNK1 KD while MMP2 expression is lost under JNK1 KD.

Article Snippet: Rabbit anti-human JNK1 and JNK2 polyclonal antibodies (sc-571 and sc-572 for JNK1 and JNK2, respectively, from Santa Cruz (Santa Cruz, CA)) were used (1:2000 and 1:1000, respectively) to detect JNK1 and JNK2 protein.

Techniques: Expressing

Journal: Carcinogenesis

Article Title: c-Jun N-terminal kinase (JNK1) upregulates XIAP-associated factor 1 (XAF1) through interferon regulatory factor 1 (IRF-1) in gastrointestinal cancer.

doi: 10.1093/carcin/bgn271

Figure Lengend Snippet: Fig. 1. XAF1 expression was upregulated by JNK1. (A) AGS and SW1116 cells were transiently transfected with pcDNA3 vector control (vector), JNK1-WTor JNK1-DN constructs. XAF1 expression was detected by reverse transcription–PCR with GAPDH being used as the internal control. (B) XAF1 and phospho-JNK1 (p-JNK1) expressions were detected by immunoblotting using whole-cell lysate of AGS cell 48 h after the transient transfection. Actin was used as the internal control. (C) AGS cells were treated with the indicated concentration of JNK inhibitor for 48 h. XAF1 and p-JNK1 were detected by immunoblotting. (D) SW1116 cells were stimulated with the indicated concentration of PMA in absence or presence of JNK inhibitor (20 lM). The expression of XAF1 and p-JNK1 was with GAPDH and actin as the internal controls. All these figures were representative of four independent experiments with the similar findings.

Article Snippet: Goat anti-human XAF1 (C-16), rabbit anti-human JNK1 (C-17) and IRF-1 (C-20), mouse anti-human c-Jun (G-4), p-JNK (G-7) and actin (I-10) and horseradish peroxidase-conjugated anti-goat, anti-rabbit and anti-mouse IgG were purchased from Santa Cruz Abbreviations: AP-1, activator protein-1; cDNA, complementary DNA; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GI, gastrointestinal; IFN, interferon; IRF-1, interferon regulatory factor 1; IRF-E, interferon regulatory factor 1-binding element; JNK, c-Jun N-terminal kinase; PCR, polymerase chain reaction; PMA, phorbol-12myristate-13-acetate; RLU, relative luciferase units; siRNA, small-interfering RNA; TNF, tumor necrosis factor; XAF1, X-linked inhibitor of apoptosis protein-associated factor 1; XIAP, X-linked inhibitor of apoptosis protein.

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Construct, Reverse Transcription, Western Blot, Concentration Assay

Journal: Carcinogenesis

Article Title: c-Jun N-terminal kinase (JNK1) upregulates XIAP-associated factor 1 (XAF1) through interferon regulatory factor 1 (IRF-1) in gastrointestinal cancer.

doi: 10.1093/carcin/bgn271

Figure Lengend Snippet: Fig. 2. JNK activity was essential for XAF1 expression. (A) AGS cells were treated with the indicated concentration of TNF-a and IFN-a in the absence or presence of JNK inhibitor (20 lM). XAF1 expression was detected by reverse transcription–PCR. (B) AGS cells with treatment of JNK1 inhibitor (20 lM) were stimulated by TNF-a and IFN-a for 48 h. The protein expressions were detected by immunoblotting. (C) AGS cells were treated with TNF-a 24 h after the transfection of control (GL2) or JNK1 siRNA for additional 48 h. Protein expressions were detected by immunoblotting. (D) AGS cells were treated with TNF-a (100 ng/ml), IFN-a (125 U/ml) or PMA (100 lM) in absence or presence of sodium orthovanadate (0.5 mM). The expressions of XAF1 and p-JNK1 were detected. All these figures were representatives of three to four independent experiments with similar findings.

Article Snippet: Goat anti-human XAF1 (C-16), rabbit anti-human JNK1 (C-17) and IRF-1 (C-20), mouse anti-human c-Jun (G-4), p-JNK (G-7) and actin (I-10) and horseradish peroxidase-conjugated anti-goat, anti-rabbit and anti-mouse IgG were purchased from Santa Cruz Abbreviations: AP-1, activator protein-1; cDNA, complementary DNA; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GI, gastrointestinal; IFN, interferon; IRF-1, interferon regulatory factor 1; IRF-E, interferon regulatory factor 1-binding element; JNK, c-Jun N-terminal kinase; PCR, polymerase chain reaction; PMA, phorbol-12myristate-13-acetate; RLU, relative luciferase units; siRNA, small-interfering RNA; TNF, tumor necrosis factor; XAF1, X-linked inhibitor of apoptosis protein-associated factor 1; XIAP, X-linked inhibitor of apoptosis protein.

Techniques: Activity Assay, Expressing, Concentration Assay, Reverse Transcription, Western Blot, Transfection, Control

Journal: Carcinogenesis

Article Title: c-Jun N-terminal kinase (JNK1) upregulates XIAP-associated factor 1 (XAF1) through interferon regulatory factor 1 (IRF-1) in gastrointestinal cancer.

doi: 10.1093/carcin/bgn271

Figure Lengend Snippet: Fig. 3. JNK1 stimulated XAF1 expression through transcriptional regulation. (A) AGS cells were treated with TNF-a (100 ng/ml), IFN-a (125 U/ml) or PMA (100 lM) in absence or presence of emetine (20 lM) or cycloheximide (20 lg/ml). XAF1 expression was evaluated by reverse transcription–PCR and real-time PCR with GAPDH as the internal control. (B) The cells were cotransfected with pGL3 basic vector or XAF1 promoter reporter construct pLUC107 together with vector control, JNK1-WT or JNK1-DN. Dual luciferase assay was performed 48 h later. RLU 5 values of firefly luciferase unit/values of Renilla luciferase unit standardized by pGL3 basic vector. P , 0.05 versus vector control. (C) AGS and SW1116 cells were transfected with pLUC107 reporter plasmid followed by the treatment with vehicle control or SP600125 (20 lM) for 48 h. Dual luciferase was performed. P , 0.05 versus vehicle control. The dual luciferase data were expressed as mean ± SD of triplicated wells. The figures were the representative of two to three independent experiments with similar results.

Article Snippet: Goat anti-human XAF1 (C-16), rabbit anti-human JNK1 (C-17) and IRF-1 (C-20), mouse anti-human c-Jun (G-4), p-JNK (G-7) and actin (I-10) and horseradish peroxidase-conjugated anti-goat, anti-rabbit and anti-mouse IgG were purchased from Santa Cruz Abbreviations: AP-1, activator protein-1; cDNA, complementary DNA; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GI, gastrointestinal; IFN, interferon; IRF-1, interferon regulatory factor 1; IRF-E, interferon regulatory factor 1-binding element; JNK, c-Jun N-terminal kinase; PCR, polymerase chain reaction; PMA, phorbol-12myristate-13-acetate; RLU, relative luciferase units; siRNA, small-interfering RNA; TNF, tumor necrosis factor; XAF1, X-linked inhibitor of apoptosis protein-associated factor 1; XIAP, X-linked inhibitor of apoptosis protein.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Plasmid Preparation, Construct, Luciferase, Transfection

Journal: Carcinogenesis

Article Title: c-Jun N-terminal kinase (JNK1) upregulates XIAP-associated factor 1 (XAF1) through interferon regulatory factor 1 (IRF-1) in gastrointestinal cancer.

doi: 10.1093/carcin/bgn271

Figure Lengend Snippet: Fig. 4. JNK1 induced IRF-1 expression. (A) XAF1 and c-Jun expressions in AGS cells transfected with GL2 or c-Jun siRNA followed by stimulation of PMA (100 lM) for 48 h were detected by immunoblotting. (B–D) The cells were transfected with vector or JNK1-WT construct (B), treated with various stimuli (C) or SP600125 (D) for 48 h. Protein expression of IRF-1 and p-JNK1 was detected by immunoblotting with actin as the control. These figures were all representatives of two independent experiments with similar findings in both AGS and SW1116 cells.

Article Snippet: Goat anti-human XAF1 (C-16), rabbit anti-human JNK1 (C-17) and IRF-1 (C-20), mouse anti-human c-Jun (G-4), p-JNK (G-7) and actin (I-10) and horseradish peroxidase-conjugated anti-goat, anti-rabbit and anti-mouse IgG were purchased from Santa Cruz Abbreviations: AP-1, activator protein-1; cDNA, complementary DNA; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GI, gastrointestinal; IFN, interferon; IRF-1, interferon regulatory factor 1; IRF-E, interferon regulatory factor 1-binding element; JNK, c-Jun N-terminal kinase; PCR, polymerase chain reaction; PMA, phorbol-12myristate-13-acetate; RLU, relative luciferase units; siRNA, small-interfering RNA; TNF, tumor necrosis factor; XAF1, X-linked inhibitor of apoptosis protein-associated factor 1; XIAP, X-linked inhibitor of apoptosis protein.

Techniques: Expressing, Transfection, Western Blot, Plasmid Preparation, Construct, Control

Journal: Carcinogenesis

Article Title: c-Jun N-terminal kinase (JNK1) upregulates XIAP-associated factor 1 (XAF1) through interferon regulatory factor 1 (IRF-1) in gastrointestinal cancer.

doi: 10.1093/carcin/bgn271

Figure Lengend Snippet: Fig. 5. IRF-1 was required in JNK1-induced XAF1 transcription. (A) AGS cells were transfected with control siRNA or IRF-1 siRNA for 48 h. IRF-1 protein was detected by immunoblotting with actin as the control. (B) AGS and SW1116 cells were cotransfected with pLUC107 and control or IRF-1 siRNA in the absence or presence of 100 lM PMA. Dual luciferase assay was measured 48 h later. P , 0.05 comparing with the control. #P . 0.05 versus control. The dual luciferase data were mean ± SD of triplicated wells and these figures were the representative of three to four independent experiments with similar results.

Article Snippet: Goat anti-human XAF1 (C-16), rabbit anti-human JNK1 (C-17) and IRF-1 (C-20), mouse anti-human c-Jun (G-4), p-JNK (G-7) and actin (I-10) and horseradish peroxidase-conjugated anti-goat, anti-rabbit and anti-mouse IgG were purchased from Santa Cruz Abbreviations: AP-1, activator protein-1; cDNA, complementary DNA; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GI, gastrointestinal; IFN, interferon; IRF-1, interferon regulatory factor 1; IRF-E, interferon regulatory factor 1-binding element; JNK, c-Jun N-terminal kinase; PCR, polymerase chain reaction; PMA, phorbol-12myristate-13-acetate; RLU, relative luciferase units; siRNA, small-interfering RNA; TNF, tumor necrosis factor; XAF1, X-linked inhibitor of apoptosis protein-associated factor 1; XIAP, X-linked inhibitor of apoptosis protein.

Techniques: Transfection, Control, Western Blot, Luciferase

Journal: Carcinogenesis

Article Title: c-Jun N-terminal kinase (JNK1) upregulates XIAP-associated factor 1 (XAF1) through interferon regulatory factor 1 (IRF-1) in gastrointestinal cancer.

doi: 10.1093/carcin/bgn271

Figure Lengend Snippet: Fig. 6. Site mutation of IRF-E abrogated JNK1-induced XAF1 transcription. (A) List of WT and 34 site mutated IRF-E within the promoter of XAF1 gene with ATG translation starting codon defined as 0. (B and C) AGS and SW1116 cells were transfected with WT (pLUC107-WT) of 34 mutated (pLUC107-34-MT) XAF1 promoter constructs followed by the treatment of PMA (B) or SP600125 (C). Dual luciferase assay was measured 48 h later. P , 0.05 versus control. #P . 0.05 versus control. The dual luciferase data were mean ± SD of triplicated wells and these figures were the representative of three independent experiments in both AGS and SW1116 cells with similar findings.

Article Snippet: Goat anti-human XAF1 (C-16), rabbit anti-human JNK1 (C-17) and IRF-1 (C-20), mouse anti-human c-Jun (G-4), p-JNK (G-7) and actin (I-10) and horseradish peroxidase-conjugated anti-goat, anti-rabbit and anti-mouse IgG were purchased from Santa Cruz Abbreviations: AP-1, activator protein-1; cDNA, complementary DNA; ERK, extracellular signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GI, gastrointestinal; IFN, interferon; IRF-1, interferon regulatory factor 1; IRF-E, interferon regulatory factor 1-binding element; JNK, c-Jun N-terminal kinase; PCR, polymerase chain reaction; PMA, phorbol-12myristate-13-acetate; RLU, relative luciferase units; siRNA, small-interfering RNA; TNF, tumor necrosis factor; XAF1, X-linked inhibitor of apoptosis protein-associated factor 1; XIAP, X-linked inhibitor of apoptosis protein.

Techniques: Mutagenesis, Transfection, Construct, Luciferase, Control